USOO94995.74B2

(12) United States Patent (10) Patent No.: US 9,499,574 B2 Miodragovic et al. (45) Date of Patent: Nov. 22, 2016

(54) ARSENOPLATIN ANTI-CANCERAGENTS OTHER PUBLICATIONS (71) Applicant: Northwestern University, Evanston, IL Miodragovic et al. Angewandte Chemie, International Edition (US) (2013), 52(41), 10749-10752.* International Search Report dated Nov. 13, 2013. (72) Inventors: Denana U. Miodragovic, Chicago, IL Anderson et al., “An expanded genetic code with a functional (S. home V. O'Halloran,9 Chicago, quadruplet7571 (2004). codon.” Proc. Natl. Acad. Sci. U.S.A. 101 (20):7566 Bacher et al., “Selection and Characterization of Escherichia coli Variants Capable of Growth on an Otherwise Toxic Trvptophan (73) Assignee: Northwestern University, Evanston, IL Analogue.” Bacteriol. 183(18):5414-5425 (2001). ryptop (US) Budisa et al., “Proteins with (beta)-(thienopyrrolyl)alanines as alter native chromophores and pharmaceutically active amino acids.” (*) Notice: Subject to any disclaimer, the term of this Protein Sci. 10(7): 1281-1292 (2001). patent is extended or adjusted under 35 Chin et al., “An Expanded Eukaryotic Genetic Code.” Science U.S.C. 154(b) by 0 days. 301(5635):964-967 (2003). Hamano-Takaku et al., “A Mutant Escherichia coli Tyrosyl-tRNA (21) Appl. No.: 14/421,982 Synthetase Utilizes the Unnatural Amino Acid AZatyrosine More Efficiently than Tyrosine,” J. Biol. Chem. 275(51):40324-40328 (22) PCT Filed: Aug. 14, 2013 (2000). Ibba et al., “Genetic code: introducing pyrrolysine.” Curr Biol. 12(13):R464-R466 (2002). (86). PCT No.: PCT/US2013/0549.99 Ikeda et al., “Synthesis of a novel histidine analogue and its efficient S 371 (c)(1), incorporation into a protein in vivo..” Protein Eng. Des. Sel. (2) Date: Feb. 16, 2015 16(9):699-706 (2003). James et al., “Kinetic characterization of ribonuclease S mutants (87) PCT Pub. No.: WO2014/028653 containing photoisomerizable phenylaZophenylalanine residues.” Protein Eng. Des. Sel. 14(12):983-991 (2001). PCT Pub. Date: Feb. 20, 2014 Stadtman, “Selenocysteine.” Annu Rev Biochem. 65:83-100 (1996). (65) Prior Publication Data Stryer et al., Biochemistry, 5. Sup,th ed., Freeman and Company (2002). US 2015/0218195 A1 Aug. 6, 2015 Zhang et al., “Selective incorporation of 5-hydroxytryptophan into O O proteins in mammalian cells.” Proc. Natl. Acad. Sci. U.S.A. Related U.S. Application Data 101(24):8882-8887 (2004). (60) Provisional application No. 61/683,031, filed on Aug. * cited by examiner 14, 2012. (51) Soon (2006.01) Primary Examiner — Sudhakar Katakam C07F 15/00 (2006.01) F.74). 2. Att Orney, AgentAgent, or Firm - Klintworth1WO & ROZa blat C07F 15/04 (2006.01) C07F 9/90 (2006.01) C07F 9/94 (2006.01) (57) ABSTRACT C07F 9/70 (2006.01) (52) U.S. Cl. Disclosed are compound having the structure of formula (I): CPC ...... C07F 19/005 (2013.01); C07F 9/70 (2013.01); C07F 9/902 (2013.01); C07F 9/94 (2013.01); C07F 15/0013 (2013.01); C07F X (I) 15/0066 (2013.01); C07F 15/045 (2013.01) (58) Field of Classification Search Al-y- & CPC ...... C07F 19/005; CO7F 9/70; C07F 9/902; L w L2 C07F 9/94; C07F 15/045; CO7F 15/0013; \,, g s CO7F 15/0066 Z -j-Z See application file for complete search history. Yi Y2 (56) References Cited wherein M is Pt, Pd or Ni; Q is As, Sb or Bi; Z' is N: Z is U.S. PATENT DOCUMENTS O or S; L and L are independently C(O), C R' or C R: X is a Lewis base and Y and Y are independently oxygen 2004/0258759 A1 12, 2004 SuSlick et al. or Sulfur-containing Substituents, conjugates containing FOREIGN PATENT DOCUMENTS compounds of formula (I) and their use as chemotherapy agents. CA 1196004 10, 1985 CN 1.01434622 5, 2009 EP O306605 3, 1989 12 Claims, 8 Drawing Sheets U.S. Patent Nov. 22, 2016 Sheet 1 of 8 US 9,499,574 B2

C1

FIG. 1A U.S. Patent Nov. 22, 2016 Sheet 2 of 8 US 9,499,574 B2

FIG. B U.S. Patent Nov. 22, 2016 Sheet 3 of 8 US 9,499,574 B2

O O-2cs Y5

-& C

FIG. 1C U.S. Patent Nov. 22, 2016 Sheet 4 of 8 US 9,499,574 B2

FIG. 1D U.S. Patent Nov. 22, 2016 Sheet S of 8 US 9,499,574 B2

H 2.18(ppm) 2.14 2.10

Nois & Sois & -3724 ppm -386 ppm Jaspesin a 456 Hz

-3710 -3750 -3790 -3830 -38to

FIG 2 U.S. Patent Nov. 22, 2016 Sheet 6 of 8 US 9,499,574 B2

A2780CP K Complex 1 00 e Cisplatin & Arsenic trioxide

50

haarara Control.0 5 2.0 2.5 3.0 LogM/u M

HTC 116

K Complex 1 e Cisplatin & Arsenic trioxide

LogM/LM FG. 3 U.S. Patent Nov. 22, 2016 Sheet 7 of 8 US 9,499,574 B2

U87 a Complex 1 9 Cisplatin 00- s : x & Arsenic trioxide

50

O NS i. orrooroo LogM/u M

D A2780 50 Complex 1 0 Cisplatin

C & Arsenic trioxide ass 00 -k-k s XX 98. xA as a O) as N. N. St 50 O 8

On ...... aga i. hoovoo Control O 2 3 LogM/u M

FIG. 3 (Cont.) U.S. Patent Nov. 22, 2016 Sheet 8 of 8 US 9,499,574 B2

E MDA-MB-231-mCherry

150 Complex 1 e Cisplatin 100 is is S- a Arsenic trioxide SS58 l r

re. Control - O 2 3 LogM/M F RPM 8226 150 Complex 1 C e Cisplatin is 100 = -s & Arsenic trioxide 9 & X O) as a teen

O

Control -1 O 2 3 LogM/u M

FIG. 3 (Cont.) US 9,499,574 B2 1. 2 ARSENOPLATIN ANT-CANCERAGENTS X (I) CROSS-REFERENCE TO RELATED APPLICATIONS A.z-- y The present application is the National Stage of Interna Z2-Q-Z2 tional Application No. PCT/US2013/054999, filed Aug. 14, M. v. 2013, and entitled “ARSENOPLATIN ANTI-CANCER Yi Y2 AGENTS, which claims the benefit of U.S. Provisional 10 Application No. 61/683,031 filed Aug. 14, 2012, and entitled wherein M is Pt, Pd or Ni; Q is As, Sb or Bi; Z' is N: Z is ARSENOPLATINS-A NEW CLASS OF ANTI-CANCER O or S; L and L are independently C(O), C R' or C R: AGENTS. The content of the U.S. Provisional Patent X is a Lewis base; Y' and Y are independently selected Application is hereby incorporated by reference in its from —OR. -OR', SR and SR, wherein R' and Rare entirety. 15 independently selected from hydrogen, halogen, cyano, keto, ester, ether, thiol, thioether, thioester, imino, C-Co STATEMENT REGARDING FEDERALLY alkyl, C-Co alkenyl, alkynyl, alkoxyl, amino, amidyl, SPONSORED RESEARCH OR DEVELOPMENT immino, Sulfonyl, Sulfoxyl, phosphoryl, phosphoryl ester, glycosyl, aryl, C-C cycloalkyl, heteroaryl, and C-Cls This invention was made with government Support under heterocycloalkyl: grant numbers U01 CA151461, U54 CA119341, U54 wherein R and Rare independently selected from hydro CA143869, P50 CA090386 awarded by the National Insti gen, C-C alkyl, C-Coalkenyl, alkynyl, alkoxyl, amino, tutes of Health and grant number W81XWH-08-1-0672 amidyl, immino, Sulfonyl, Sulfoxyl, phosphoryl, phosphoryl awarded by the US Army Medical Research and Materiel ester, glycosyl, aryl, C-C cycloalkyl, heteroaryl, and Command. The government has certain rights in the inven 25 C-Cls heterocycloalkyl; tion. wherein the Lewis base can be halogen, N-bonded cyano, nitroso, nitroxyl, N-bonded C-C alkyl, C-C alkenyl, BACKGROUND N-bonded C-C cycloalkyl, N-bonded C-Coheteroalkyl, N-bonded C-Co heteroalkenyl, N-bonded aryl, N-bonded 1. Technical Field 30 heteroaryl, N-bonded C-Cls heterocycloalkyl, NR'R''. The disclosure relates to arsenoplatin and related com N-bonded ligand, S-bonded cyano, sulfonyl, sulfoxyl, thiol, pounds and methods for their use as chemotherapy agents. S-bonded thioether, S-bonded thioester, S-bonded C-Co 2. Description of Relevant Prior Art alkyl, C-Co alkenyl, S-bonded C-Co cycloalkyl, Cis-diamminedichloroplatinum(II) ("Cisplatin') and S-bonded C-Coheteroalkyl, S-bonded C-C heteroalk 35 enyl, S-bonded aryl, S-bonded heteroaryl, S-bonded C-Cls arsenic trioxide (ASOs) are highly successful agents for heterocycloalkyl, - SR, S-bonded ligand, O-bonded treatment of cancer. Cisplatin is used in combination che C-C cycloalkyl, O-bonded C-C heteroalkyl, O-bonded motherapy to treat ovarian, testicular, head, neck, and blad C-Co heteroalkenyl, O-bonded aryl, O-bonded heteroaryl, der cancers. Unfortunately, these and other cancers fre O-bonded C-Cls heterocycloalkyl, —OR, O-bonded car quently develop resistance to this widely used agent and 40 boxylato, O-bonded polycarboxylato, O-bonded carboxy there are intensive efforts to develop new agents that over lato, O-bonded polycarboxylato, O-bonded carboxylato, come this resistance. Arsenic trioxide, which was discovered O-bonded polycarboxylato, O-bonded ligand, or P-bonded as a traditional Chinese medicine, is a front line treatment phosphine having formula P(R)-(R), where x is 0, 1, 2 for acute promyelocytic leukemia and has also shown pre or 3: liminary efficacy in the treatment of blood cancers such as 45 wherein R and Rare independently selected from hydro multiple myeloma and myelodysplastic syndromes. gen, O. —CO.R. —COR, C-Clo alkyl, C-Coalkenyl, Both compounds induce apoptotic cell death, but through aryl, heteroaryl, C-C cycloalkyl and C-Cls heterocy different pathways. Cisplatin reacts with DNA and causes cloalkyl; intra- and inter-strand DNA cross-links. The principal com wherein R and Rare independently selected from C-Co ponent of aqueous Solutions of ASO at pH 7. arsenous acid, 50 alkyl, C-Co alkenyl, aryl, heteroaryl, C-C cycloalkyl at low concentrations reacts with and triggers degradation of and C-Cls heterocycloalkyl; and key zinc-dependent regulatory proteins and inhibits angio wherein R is selected from hydrogen, hydroxyl, halogen, genesis, migration and invasion. At higher concentrations, amine, thiol, ether, C-C alkyl, C-C alkenyl, aryl, het arsenous acid triggers apoptosis through pathways that eroaryl, C-C cycloalkyl and C-Cls heterocycloalkyl. involve elevated levels of reactive oxygen species in mito 55 In a second aspect, conjugate that includes a ligand and a chondria. compound having the structure of formula (I) is disclosed: When combined, these agents can act synergistically in certain cisplatin sensitive and resistant ovarian and non Small lung carcinoma cells. The only example of a platinum (I) adduct with arsenous acid in the literature emerged in efforts 60 to develop efficient systems for loading ASO into lipo Somes with aquated forms of cisplatin. BRIEF SUMMARY 65 In a first aspect, a compound having the structure of formula (I) is disclosed: US 9,499,574 B2 3 4 wherein M is Pt, Pd or Ni; Q is As, Sb or Bi; Z is N; Z is wherein R and Rare independently selected from hydro O or S: L' and L are independently C(O), C R' or C R: gen, C-C alkyl, C-Coalkenyl, alkynyl, alkoxyl, amino, X is a Lewis base; Y' and Y are independently selected amidyl, immino, Sulfonyl, Sulfoxyl, phosphoryl, phosphoryl from OR, OR, -SR and SR, wherein R' and Rare ester, glycosyl, aryl, C-C cycloalkyl, heteroaryl, and independently selected from hydrogen, halogen, cyano, C-Cls heterocycloalkyl; keto, ester, ether, thiol, thioether, thioester, imino, C-Co wherein the Lewis base can be halogen, N-bonded cyano, alkyl, C-Co alkenyl, alkynyl, alkoxyl, amino, amidyl, nitroso, nitroxyl, N-bonded C-Co alkyl, C-Co alkenyl, immino, Sulfonyl, Sulfoxyl, phosphoryl, phosphoryl ester, N-bonded C-C cycloalkyl, N-bonded C-Coheteroalkyl, glycosyl, aryl, C-C cycloalkyl, heteroaryl, and C-Cls N-bonded C-Co heteroalkenyl, N-bonded aryl, N-bonded heterocycloalkyl: 10 heteroaryl, N-bonded C-Cls heterocycloalkyl, NR'R''. wherein R and R are independently selected from hydro N-bonded ligand, S-bonded cyano, sulfonyl, sulfoxyl, thiol, gen, C-C alkyl, C-Coalkenyl, alkynyl, alkoxyl, amino, S-bonded thioether, S-bonded thioester, S-bonded C-Co amidyl, immino, Sulfonyl, Sulfoxyl, phosphoryl, phosphoryl alkyl, C-Co alkenyl, S-bonded C-Co cycloalkyl, ester, glycosyl, aryl, C-C cycloalkyl, heteroaryl, and S-bonded C-C heteroalkyl, S-bonded C-C heteroalk 15 enyl, S-bonded aryl, S-bonded heteroaryl, S-bonded C-Cls C-Cls heterocycloalkyl; heterocycloalkyl, - SR, S-bonded ligand, O-bonded wherein the Lewis base can be halogen, N-bonded cyano, C-C cycloalkyl, O-bonded C-C heteroalkyl, O-bonded nitroso, nitroxyl, N-bonded C-C alkyl, C-C alkenyl, C-Co heteroalkenyl, O-bonded aryl, O-bonded heteroaryl, N-bonded C-C cycloalkyl, N-bonded C-Coheteroalkyl, O-bonded C-Cls heterocycloalkyl, —OR, O-bonded car N-bonded C-Co heteroalkenyl, N-bonded aryl, N-bonded boxylato, O-bonded polycarboxylato, O-bonded ligand, or heteroaryl, N-bonded C-Cls heterocycloalkyl, NR'R''. P-bonded phosphine having formula P(R)-(R), where x N-bonded ligand, S-bonded cyano, sulfonyl, sulfoxyl, thiol, is 0, 1, 2 or 3: S-bonded thioether, S-bonded thioester, S-bonded C-Co wherein R and Rare independently selected from hydro alkyl, C-C alkenyl, S-bonded C-C cycloalkyl, gen, O. —CO.R. —COR, C-Clo alkyl, C-Coalkenyl, S-bonded C-C heteroalkyl, S-bonded C-C heteroalk 25 aryl, heteroaryl, C-C cycloalkyl and C-Cls heterocy enyl, S-bonded aryl, S-bonded heteroaryl, S-bonded C-C, cloalkyl; heterocycloalkyl, - SR, S-bonded ligand, O-bonded wherein R and Rare independently selected from C-Co C-C cycloalkyl, O-bonded C-Coheteroalkyl, O-bonded alkyl, C-Co alkenyl, aryl, heteroaryl, C-C cycloalkyl C-C heteroalkenyl, O-bonded aryl, O-bonded heteroaryl, and C-Cls heterocycloalkyl; and O-bonded C-Cls heterocycloalkyl, OR, O-bonded car 30 wherein R is selected from hydrogen, hydroxyl, halogen, boxylato, O-bonded polycarboxylato, O-bonded ligand, or amine, thiol, ether, C-C alkyl, C-C alkenyl, aryl, het P-bonded phosphine having formula P(R)-(R), where x eroaryl, C-Cs cycloalkyl and C-Cls heterocycloalkyl. is 0, 1, 2 or 3: In a fourth aspect, a pharmaceutical composition that wherein R and R are independently selected from hydro includes an excipient and a compound having the structure gen, O. —CO.R. —COR, C-Clo alkyl, C-Clo alkenyl, 35 of formula (I) is disclosed: aryl, heteroaryl, C-C cycloalkyl and C-Cls heterocy cloalkyl; wherein R7 and Rare independently selected from C-Co (I) alkyl, C-C alkenyl, aryl, heteroaryl, C-C cycloalkyl and C-Cls heterocycloalkyl, and 40 i wherein R is selected from hydrogen, hydroxyl, halogen, Al-Y- amine, thiol, ether, C-Co alkyl, C-Coalkenyl, aryl, het L & ty 2 eroaryl, C-C cycloalkyl and C-Cls heterocycloalkyl. Z2-Q-Z2 /v In a third aspect, a method of treating a cancer cell that Yi Y2 includes the step of contacting the cancer cell with a 45 compound having the structure of formula (I) is disclosed: wherein M is Pt, Pd or Ni; Q is As, Sb or Bi; Z' is N: Z is O or S; L and L are independently C(O), C R' or C R: (I) X is a Lewis base; Y' and Y are independently selected 50 from —OR. -OR. -SR and - SR", wherein R' and R' z-- are independently selected from hydrogen, halogen, cyano, A. y keto, ester, ether, thiol, thioether, thioester, imino, C-Co ', L. alkyl, C-Co alkenyl, alkynyl, alkoxyl, amino, amidyl, z?--z immino, Sulfonyl, Sulfoxyl, phosphoryl, phosphoryl ester, M. v. 55 glycosyl, aryl, C-C cycloalkyl, heteroaryl, and C-Cls Yi Y2 heterocycloalkyl: wherein R and Rare independently selected from hydro wherein M is Pt, Pd or Ni; Q is As, Sb or Bi; Z is N; Z is gen, C-C alkyl, C-Coalkenyl, alkynyl, alkoxyl, amino, O or S: L' and L are independently C(O), C R' or C R: amidyl, immino, Sulfonyl, Sulfoxyl, phosphoryl, phosphoryl X is a Lewis base; Y and Y are independently selected 60 ester, glycosyl, aryl, C-C cycloalkyl, heteroaryl, and from -OR. -OR. -SR and - SR", wherein R' and R' C-Cls heterocycloalkyl; are independently selected from hydrogen, halogen, cyano, wherein the Lewis base can be halogen, N-bonded cyano, keto, ester, ether, thiol, thioether, thioester, imino, C-Co nitroso, nitroxyl, N-bonded C-C alkyl, C-C alkenyl, alkyl, C-Co alkenyl, alkynyl, alkoxyl, amino, amidyl, N-bonded C-C cycloalkyl, N-bonded C-Coheteroalkyl, immino, Sulfonyl, Sulfoxyl, phosphoryl, phosphoryl ester, 65 N-bonded C-Co heteroalkenyl, N-bonded aryl, N-bonded glycosyl, aryl, C-C cycloalkyl, heteroaryl, and C-Cls heteroaryl, N-bonded C-Cls heterocycloalkyl, NR'R''. heterocycloalkyl: N-bonded ligand, S-bonded cyano, sulfonyl, sulfoxyl, thiol, US 9,499,574 B2 5 6 S-bonded thioether, S-bonded thioester, S-bonded C-Co FIG. 3F depicts a dose response curve for cell growth alkyl, C-C alkenyl, S-bonded C-C cycloalkyl, inhibition of multiple myeloma RPMI 8226 cells following S-bonded C-C heteroalkyl, S-bonded C-C heteroalk exposure to one of compound (1), cisplatin or ASO. enyl, S-bonded aryl, S-bonded heteroaryl, S-bonded C-Cls heterocycloalkyl, - SR, S-bonded ligand, O-bonded DETAILED DESCRIPTION C-C cycloalkyl, O-bonded C-Coheteroalkyl, O-bonded C-Co heteroalkenyl, O-bonded aryl, O-bonded heteroaryl, In the following detailed description, reference is made to O-bonded C-Cls heterocycloalkyl, OR, O-bonded car the accompanying figures, which form a part hereof. In the boxylato, O-bonded polycarboxylato, O-bonded ligand, or figures, similar symbols typically identify similar compo P-bonded phosphine having formula P(R)-(R), where x 10 nents, unless context dictates otherwise. Insofar as possible, is 0, 1, 2 or 3: like parts and modules have the same reference numeral in wherein R and Rare independently selected from hydro the figures. The illustrative embodiments described in the gen, O. —CO.R. —COR, C-Clo alkyl, C-Clo alkenyl, detailed description, figures, and claims are not meant to be aryl, heteroaryl, C-C cycloalkyl and C-Cls heterocy limiting. Other embodiments may be utilized, and other 15 changes may be made, without departing from the spirit or cloalkyl; scope of the subject matter presented herein. It will be wherein R and Rare independently selected from C-Co readily understood that the aspects of the present disclosure, alkyl, C-C alkenyl, aryl, heteroaryl, C-C cycloalkyl as generally described herein, and illustrated in the figures, and C-Cls heterocycloalkyl, and can be arranged, Substituted, combined, separated, and wherein R is selected from hydrogen, hydroxyl, halogen, designed in a wide variety of different configurations, all of amine, thiol, ether, C-C alkyl, C-C alkenyl, aryl, het which are contemplated herein. eroaryl, C-C cycloalkyl and C-Cls heterocycloalkyl. The present disclosure pertains to synthesis and charac terization of a novel family of Small molecule complexes, BRIEF DESCRIPTION OF THE DRAWINGS such as an aqueous form of AsO. bound directly to Pt as 25 an As(OH), moiety, wherein an As' center acts simultane FIG. 1A depicts a thermal ellipsoid plot of compound (1), ously as a Lewis acid and a Lewis base. These arsenoplatins as crystallized in a triclinic form (solvent molecules omitted and related compounds are Surprisingly stable in Solution for clarity) with space group P-1. The plot is shown at 50% and exhibit unexpected chemical bonding, novel ligand probability level. Substitution chemistry, and biological activities that are FIG. 1B depicts a thermal ellipsoid plot of compound (1), 30 distinct from the parent compounds. The class of arsenopla as crystallized in a monoclinic form with lattice acetamide tins and related compounds demonstrate promising activity molecules (solvent molecules omitted for clarity) with space as chemotherapeutic agent in drug-resistant cancer cells. group P2(1)/n. The plot is shown at 50% probability level. One embodiment pertains to a compound having formula FIG. 1C depicts a thermal ellipsoid plot of compound (2) (I): (solvent molecules omitted for clarity). The plot is shown at 35 50% probability level. FIG. 1D depicts a thermal ellipsoid plot of compound (5) (I) (solvent molecules omitted for clarity). The plot is shown at 50% probability level. z-- FIG. 2A depicts a 'H-'C HSQC NMR spectrum of 40 2 compound (5) (with S'C'N) in DDMSO, acquired at 25° C. at 600 MHz H with high resolution in the indirect (C) dimension to distinguish the methyl resonances of the N- and S-isomers. 45 FIG. 2B depicts a 'Pt NMR spectrum of compound (5) wherein M is Pt, Pd or Ni; Q is As, Sb or Bi; Z' is N; Z is (with S'C'N) in DDMSO, referenced indirectly to H O or S; L and L are independently C(O), C R' or C R: TMS such that Na'PtCl resonates at 0.0 ppm. The 456 X is a Lewis base; Y and Y are independently selected Hz splitting of the 'Pt peak at -3724 ppm arises from from -OR. -OR. -SR and - SR", wherein R' and R' scalar coupling to the SCNN. 50 are independently selected from hydrogen, halogen, cyano, FIG. 3A depicts a dose response curve for cell growth keto, ester, ether, thiol, thioether, thioester, imino, C-Co inhibition of ovarian cisplatin-resistant cells A2780 fol alkyl, C-Co alkenyl, alkynyl, alkoxyl, amino, amidyl, lowing exposure to one of compound (1), cisplatin or ASO. immino, Sulfonyl, Sulfoxyl, phosphoryl, phosphoryl ester, FIG. 3B depicts a dose response curve for cell growth glycosyl, aryl, C-C cycloalkyl, heteroaryl, and C-Cls 55 heterocycloalkyl: inhibition of colon HCT-116 cells following exposure to one wherein R and R are independently selected from hydro of compound (1), cisplatin or ASO. gen, C-C alkyl, C-Coalkenyl, alkynyl, alkoxyl, amino, FIG. 3C depicts a dose response curve for cell growth amidyl, immino, Sulfonyl, Sulfoxyl, phosphoryl, phosphoryl inhibition of glioblastoma U-87 cells following exposure to ester, glycosyl, aryl, C-C cycloalkyl, heteroaryl, and one of compound (1), cisplatin or ASO. 60 C-Cls heterocycloalkyl; FIG. 3D depicts a dose response curve for cell growth wherein the Lewis base can be halogen, N-bonded cyano, inhibition of ovarian cisplatin-sensitive cells A2780 cells nitroso, nitroxyl, N-bonded C-Co alkyl, C-Co alkenyl, following exposure to one of compound (1), cisplatin or N-bonded C-C cycloalkyl, N-bonded C-Coheteroalkyl, ASOs. N-bonded C-Co heteroalkenyl, N-bonded aryl, N-bonded FIG. 3E depicts a dose response curve for cell growth 65 heteroaryl, N-bonded C-Cls heterocycloalkyl, NR'R''. inhibition of triple-negative breast MDA-MB-231 cells fol N-bonded ligand, S-bonded cyano, sulfonyl, sulfoxyl, thiol, lowing exposure to one of compound (1), cisplatin or ASOs. S-bonded thioether, S-bonded thioester, S-bonded C-Co US 9,499,574 B2 7 8 alkyl, C-Co alkenyl, S-bonded C-Co cycloalkyl, wherein R and Rare independently selected from C-Co S-bonded C-C heteroalkyl, S-bonded C-C heteroalk alkyl, C-C alkenyl, aryl, heteroaryl, C-C cycloalkyl enyl, S-bonded aryl, S-bonded heteroaryl, S-bonded C-Cls and C-Cls heterocycloalkyl; and heterocycloalkyl, - SR, S-bonded ligand, O-bonded wherein R is selected from hydrogen, hydroxyl, halogen, C-C cycloalkyl, O-bonded C-C heteroalkyl, O-bonded amine, thiol, ether, C-C alkyl, C-C alkenyl, aryl, het C-Co heteroalkenyl, O-bonded aryl, O-bonded heteroaryl, eroaryl, C-C cycloalkyl and C-Cls heterocycloalkyl. O-bonded C-Cls heterocycloalkyl, OR, O-bonded car Another embodiment pertains to a compound having boxylato, O-bonded polycarboxylato, O-bonded ligand, or formula (III): P-bonded phosphine having formula P(R)-(R), where x is 0, 1, 2 or 3: 10 wherein R and Rare independently selected from hydro (III) gen, O. —CO.R. —COR, C-Clo alkyl, C-Clo alkenyl, aryl, heteroaryl, C-C cycloalkyl and C-Cls heterocy Z-Pt-Zi cloalkyl; A. M L. L2, wherein R7 and Rare independently selected from C-Co 15 V, 7 alkyl, C-C alkenyl, aryl, heteroaryl, C-C cycloalkyl Z-As-Z2 and C-Cls heterocycloalkyl, and M. V. wherein R is selected from hydrogen, hydroxyl, halogen, HO OH amine, thiol, ether, C-Co alkyl, C-Coalkenyl, aryl, het wherein Z' is N; Z is O or S: L' and L are independently eroaryl, C-C cycloalkyl and C-Cls heterocycloalkyl. C(O), C R' or C R. X is a Lewis base: Another embodiment pertains to a compound having wherein R' and R are independently selected from hydro formula (II): gen, halogen, cyano, keto, ester, ether, thiol, thioether, thioester, imino, C-Co alkyl, C-Co alkenyl, alkynyl, 25 alkoxyl, amino, amidyl, immino, Sulfonyl, Sulfoxyl, phos (II) phoryl, phosphoryl ester, glycosyl, aryl, C-C cycloalkyl, heteroaryl, and C-Cls heterocycloalkyl; wherein the Lewis base can be halogen, N-bonded cyano, nitroso, nitroxyl, N-bonded C-Co alkyl, C-Co alkenyl, 30 N-bonded C-C cycloalkyl, N-bonded C-Coheteroalkyl, N-bonded C-Co heteroalkenyl, N-bonded awl, N-bonded heteroaryl, N-bonded C-C heterocycloalkyl, NRR, N-bonded ligand, S-bonded cyano, sulfonyl, sulfoxyl, thiol, wherein M is Pt, Pd or Ni; Q is As, Sb or Bi: X is a Lewis S-bonded thioether, S-bonded thioester, S-bonded C-Co base; 35 alkyl, C-C alkenyl, S-bonded C-C cycloalkyl, wherein R' and R are independently selected from hydro S-bonded C-C heteroalkyl, S-bonded C-C heteroalk gen, halogen, cyano, keto, ester, ether, thiol, thioether, enyl, S-bonded awl, S-bonded heteroaryl, S-bonded C-C, thioester, imino, C-Co alkyl, C-Co alkenyl, alkynyl, heterocycloalkyl, - SR, S-bonded ligand, O-bonded alkoxyl, amino, amidyl, immino, Sulfonyl, Sulfoxyl, phos C-C cycloalkyl, O-bonded C-C heteroalkyl, O-bonded phoryl, phosphoryl ester, glycosyl, awl, C-C cycloalkyl, 40 C-Co heteroalkenyl, O-bonded awl, O-bonded heteroaryl, heteroaryl, and C-Cls heterocycloalkyl; O-bonded C-Cls heterocycloalkyl, —OR, O-bonded car wherein R and R are independently selected from hydro boxylato, O-bonded polycarboxylato, O-bonded ligand, or gen, C-C alkyl, C-Coalkenyl, alkynyl, alkoxyl, amino, P-bonded phosphine having formula P(R)-(R), where x amidyl, immino, Sulfonyl, Sulfoxyl, phosphoryl, phosphoryl is 0, 1, 2 or 3: ester, glycosyl, aryl, C-C cycloalkyl, heteroaryl and 45 wherein R and Rare independently selected from hydro C-Cls heterocycloalkyl; gen, O. —CO.R. —COR, C-Clo alkyl, C-Clo alkenyl, wherein the Lewis base can be halogen, N-bonded cyano, aryl, heteroaryl, C-C cycloalkyl and C-Cls heterocy nitroso, nitroxyl, N-bonded C-C alkyl, C-Co alkenyl, cloalkyl; N-bonded C-C cycloalkyl, N-bonded C-C heteroalkyl, wherein R7 and Rare independently selected from C-Co N-bonded C-Co heteroalkenyl, N-bonded awl, N-bonded 50 alkyl, C-Co alkenyl, aryl, heteroaryl, C-C cycloalkyl heteroaryl, N-bonded C-Cls heterocycloalkyl, NR'R''. and C-Cls heterocycloalkyl; and N-bonded ligand, S-bonded cyano, sulfonyl, sulfoxyl, thiol, wherein R is selected from hydrogen, hydroxyl, halogen, S-bonded thioether, S-bonded thioester, S-bonded C-Co amine, thiol, ether, C-Co alkyl, C-Coalkenyl, aryl, het alkyl, C-C alkenyl, S-bonded C-C cycloalkyl, eroaryl, C-C cycloalkyl and C-Cls heterocycloalkyl. S-bonded C-C heteroalkyl, S-bonded C-C heteroalk 55 Another embodiment pertains to a compound having enyl, S-bonded awl, S-bonded heteroaryl, S-bonded C-C, formula (IV): heterocycloalkyl, - SR, S-bonded ligand, O-bonded C-C cycloalkyl, O-bonded C-Coheteroalkyl, O-bonded C-Co heteroalkenyl, O-bonded awl, O-bonded heteroaryl, (IV) O-bonded C-Cls heterocycloalkyl, OR, O-bonded car 60 boxylato, O-bonded polycarboxylato, O-bonded ligand, or P-bonded phosphine having formula P(R)-(R), where x is 0, 1, 2 or 3: wherein R and Rare independently selected from hydro gen, O. —CO.R. —COR, C-Clo alkyl, C-Clo alkenyl, 65 aryl, heteroaryl, C-C cycloalkyl and C-Cls heterocy cloalkyl; US 9,499,574 B2 9 10 wherein X is a Lewis base; wherein the Lewis base can be halogen, N-bonded cyano, wherein R' and R are independently selected from hydro nitroso, nitroxyl, N-bonded C-C alkyl, C-C alkenyl, gen, halogen, cyano, keto, ester, ether, thiol, thioether, N-bonded C-C cycloalkyl, N-bonded C-Coheteroalkyl, thioester, imino, C-Co alkyl, C-Co alkenyl, alkynyl, N-bonded C-Co heteroalkenyl, N-bonded awl, N-bonded alkoxyl, amino, amidyl, immino, Sulfonyl, Sulfoxyl, phos heteroaryl, N-bonded C-Cls heterocycloalkyl, NR'R''. phoryl, phosphoryl ester, glycosyl, aryl, C-C cycloalkyl, N-bonded ligand, S-bonded cyano, sulfonyl, sulfoxyl, thiol, heteroaryl, and C-Cls heterocycloalkyl; S-bonded thioether, S-bonded thioester, S-bonded C-Co wherein Y and Y are independently selected from —OR, alkyl, C-C alkenyl, S-bonded C-C cycloalkyl, OR', SR and SR, S-bonded C-C heteroalkyl, S-bonded C-C heteroalk wherein R and R are independently selected from hydro 10 enyl, S-bonded awl, S-bonded heteroaryl, S-bonded C-Cls gen, C-C alkyl, C-Coalkenyl, alkynyl, alkoxyl, amino, heterocycloalkyl, - SR, S-bonded ligand, O-bonded amidyl, immino, Sulfonyl, Sulfoxyl, phosphoryl, phosphoryl C-C cycloalkyl, O-bonded C-Coheteroalkyl, O-bonded ester, glycosyl, aryl, C-C cycloalkyl, heteroaryl, and C-Co heteroalkenyl, O-bonded awl, O-bonded heteroaryl, C-Cls heterocycloalkyl; 15 O-bonded C-Cls heterocycloalkyl, —OR, O-bonded car wherein the Lewis base can be halogen, N-bonded cyano, boxylato, O-bonded polycarboxylato, O-bonded ligand, or nitroso, nitroxyl, N-bonded C-C alkyl, C-Co alkenyl, P-bonded phosphine having formula P(R)-(R), where x N-bonded C-C cycloalkyl, N-bonded C-C heteroalkyl, is 0, 1, 2 or 3: N-bonded C-Co heteroalkenyl, N-bonded awl, N-bonded wherein R and Rare independently selected from hydro heteroaryl, N-bonded C-Cls heterocycloalkyl, NR'R''. gen, O. —CO.R. —COR, C-Clo alkyl, C-Clo alkenyl, N-bonded ligand, S-bonded cyano, sulfonyl, sulfoxyl, thiol, aryl, heteroaryl, C-C cycloalkyl and C-Cls heterocy S-bonded thioether, S-bonded thioester, S-bonded C-Co cloalkyl; alkyl, C-Co alkenyl, S-bonded C-Co cycloalkyl, wherein R and Rare independently selected from C-Co S-bonded C-C heteroalkyl, S-bonded C-C heteroalk alkyl, C-C alkenyl, aryl, heteroaryl, C-C cycloalkyl enyl, S-bonded awl, S-bonded heteroaryl, S-bonded C-Cls 25 and C-Cls heterocycloalkyl; and heterocycloalkyl, - SR, S-bonded ligand, O-bonded wherein R is selected from hydrogen, hydroxyl, halogen, C-C cycloalkyl, O-bonded C-Coheteroalkyl, O-bonded amine, thiol, ether, C-Co alkyl, C-Coalkenyl, aryl, het C-Co heteroalkenyl, O-bonded awl, O-bonded heteroaryl, eroaryl, C-C cycloalkyl and C-Cls heterocycloalkyl. O-bonded C-Cls heterocycloalkyl, OR, O-bonded car Another embodiment pertains to a compound having boxylato, O-bonded polycarboxylato, O-bonded ligand, or 30 formula (VI): P-bonded phosphine having formula P(R)-(R), where x is 0, 1, 2 or 3: wherein R and Rare independently selected from hydro (VI) gen, O. —CO.R. —COR, C-Clo alkyl, C-Coalkenyl, aryl, heteroaryl, C-C cycloalkyl and C-Cls heterocy 35 cloalkyl; wherein R and Rare independently selected from C-Co alkyl, C-C alkenyl, aryl, heteroaryl, C-C cycloalkyl and C-Cls heterocycloalkyl, and wherein R is selected from hydrogen, hydroxyl, halogen, 40 amine, thiol, ether, C-C alkyl, C-C alkenyl, aryl, het eroaryl, C-C cycloalkyl and C-Cls heterocycloalkyl. wherein M is Pt, Pd or Ni; Q is As, Sb or Bi: X is a Lewis Another embodiment pertains to a compound having base; formula (V): wherein R' and R are independently selected from hydro 45 gen, halogen, cyano, keto, ester, ether, thiol, thioether, thioester, imino, C-C alkyl, C-C alkenyl, alkynyl. (V) alkoxyl, amino, amidyl, immino, Sulfonyl, Sulfoxyl, phos phoryl, phosphoryl ester, glycosyl, aryl, C-C cycloalkyl, heteroaryl, and C-Cls heterocycloalkyl; and 50 wherein the Lewis base can be halogen, N-bonded cyano, nitroso, nitroxyl, N-bonded C-C alkyl, C-C alkenyl, N-bonded C-C cycloalkyl, N-bonded C-Coheteroalkyl, N-bonded C-Co heteroalkenyl, N-bonded awl, N-bonded 55 heteroaryl, N-bonded C-Cls heterocycloalkyl, NR'R''. wherein X is a Lewis base; N-bonded ligand, S-bonded cyano, sulfonyl, sulfoxyl, thiol, wherein R' and R are independently selected from hydro S-bonded thioether, S-bonded thioester, S-bonded C-Co gen, halogen, cyano, keto, ester, ether, thiol, thioether, alkyl, C-C alkenyl, S-bonded C-C cycloalkyl, thioester, imino, C-Co alkyl, C-Co alkenyl, alkynyl, S-bonded C-C heteroalkyl, S-bonded C-C heteroalk alkoxyl, amino, amidyl, immino, Sulfonyl, Sulfoxyl, phos 60 enyl, S-bonded awl, S-bonded heteroaryl, S-bonded C-Cls phoryl, phosphoryl ester, glycosyl, aryl, C-C cycloalkyl, heterocycloalkyl, - SR, S-bonded ligand, O-bonded heteroaryl, and C-Cls heterocycloalkyl; C-C cycloalkyl, O-bonded C-Coheteroalkyl, O-bonded wherein R and R are independently selected from hydro C-C heteroalkenyl, O-bonded awl, O-bonded heteroaryl, gen, C-C alkyl, C-Coalkenyl, alkynyl, alkoxyl, amino, O-bonded C-Cls heterocycloalkyl, —OR, O-bonded car amidyl, immino, Sulfonyl, Sulfoxyl, phosphoryl, phosphoryl 65 boxylato, O-bonded polycarboxylato, O-bonded ligand, or ester, glycosyl, aryl, C-C cycloalkyl, heteroaryl, and P-bonded phosphine having formula P(R)-(R), where x C-Cls heterocycloalkyl; is 0, 1, 2 or 3: US 9,499,574 B2 11 12 wherein R and Rare independently selected from hydro eties. Likewise, the cited heteroaryl and heterocycloalkyl gen, O. —CO.R. —COR, C-Clo alkyl, C-Coalkenyl, substituents include at least one heteroatom different from awl, heteroaryl, C-C cycloalkyl and C-Cls heterocy carbon (C) as part of the ring system, such as nitrogen (N), cloalkyl; oxygen (O), sulfur (S), phosphorous (P) or boron (B). Where wherein R7 and Rare independently selected from C-Co substituents include lone electron pairs available for bonding alkyl, C-C alkenyl, aryl, heteroaryl, C-C cycloalkyl to other atoms, those substituents can include further sub and C-Cls heterocycloalkyl, and stitutions. Examples of moieties that can include further wherein R is selected from hydrogen, hydroxyl, halogen, substitutions include keto, ester, ether, thioether, thioester, amine, thiol, ether, C-Co alkyl, C-Coalkenyl, aryl, het imino, amino, amidyl, immino, Sulfonyl, Sulfoxyl, phospho eroaryl, C-C cycloalkyl and C-Cls heterocycloalkyl. 10 ryl, phosphoryl ester, glycosyl and the like. Exemplary Another embodiment pertains to a compound having halogen Substituents include —F. —Cl. —Brand —I. formula (VII): As used herein, "N-bonded’ refers to a lone pair of electrons from nitrogen being coordinated to M, where M is Pt, Pd or Ni. Where "N-bonded” modifies “alkyl or “aryl.” (VII) 15 the M-coordinating nitrogen forms a bond to the respective alkyl or aryl, wherein M is Pt, Pd or Ni. An exemplary "N-bonded alkyl moiety is a methylamine radical. An exemplary "N-bonded aryl moiety is an aniline radical. Where "N-bonded” modifies a heteroatom-containing sub stituent, such as "heterocycloalkyl or "heteroaryl, the M-coordinating nitrogen forms part of the ring system of the respective heterocycloalkyl or heteroaryl, wherein M is Pt, wherein X is a Lewis base; Pd or Ni. An exemplary "N-bonded heterocycloalkyl is a wherein R' and R are independently selected from hydro 25 morpholine radical. An exemplary "N-bonded heteroaryl' is gen, halogen, cyano, keto, ester, ether, thiol, thioether, a pyridine radical. As used herein, "N-bonded ligand” refers thioester, imino, C-Co alkyl, C-Co alkenyl, alkynyl, to an M-coordinating nitrogen forming a bond to a ligand, alkoxyl, amino, amidyl, immino, Sulfonyl, Sulfoxyl, phos wherein M is Pt, Pd or Ni and ligand includes an amino acid, phoryl, phosphoryl ester, glycosyl, aryl, C-C cycloalkyl, a peptide, a protein, a glycan, a peptidoglycan, a polysac heteroaryl, and C-Cls heterocycloalkyl; and 30 charide, a biologically-compatible polymer (for example, a wherein the Lewis base can be halogen, N-bonded cyano, polyethylene glycol (PEG) polymer) or an oligonucleotide. nitroso, nitroxyl, N-bonded C-C alkyl, C-C alkenyl, As used herein, “S-bonded” refers to a lone pair of N-bonded C-C cycloalkyl, N-bonded C-C heteroalkyl, electrons from sulfur being coordinated to M, where M is Pt, N-bonded C-Co heteroalkenyl, N-bonded aryl, N-bonded Pd or Ni. Where “S-bonded” modifies “alkyl or “awl,” the heteroaryl, N-bonded C-Cls heterocycloalkyl, NR'R''. 35 M-coordinating sulfur forms a bond to the respective alkyl N-bonded ligand, S-bonded cyano, sulfonyl, sulfoxyl, thiol, or aryl, wherein Mis Pt, Pd or Ni. An exemplary “S-bonded S-bonded thioether, S-bonded thioester, S-bonded C-Co alkyl moiety is a methanethiol radical. An exemplary alkyl, C-Co alkenyl, S-bonded C-Co cycloalkyl, “S-bonded aryl moiety is a thiophenol radical. Where S-bonded C-C heteroalkyl, S-bonded C-C heteroalk “S-bonded modifies a heteroatom-containing substituent, enyl, S-bonded aryl, S-bonded heteroaryl, S-bonded C-C, 40 such as "heterocycloalkyl or "heteroaryl, the M-coordi heterocycloalkyl, - SR, S-bonded ligand, O-bonded nating Sulfur forms part of the ring system of the respective C-C cycloalkyl, O-bonded C-C heteroalkyl, O-bonded heterocycloalkyl or heteroaryl, wherein Mis Pt, Pd or Ni. An C-Co heteroalkenyl, O-bonded awl, O-bonded heteroaryl, exemplary “S-bonded heterocycloalkyl is a thioxane radi O-bonded C-Cls heterocycloalkyl, OR, O-bonded car cal. An exemplary “S-bonded heteroaryl is a thiophene boxylato, O-bonded polycarboxylato, O-bonded ligand, or 45 radical. As used herein, “S-bonded ligand” refers to an P-bonded phosphine having formula P(R)-(R), where x M-coordinating Sulfur forming a bond to a ligand, wherein is 0, 1, 2 or 3: M is Pt, Pd or Ni and ligand includes an amino acid, a wherein R and Rare independently selected from hydro peptide, a protein, a glycan, a peptidoglycan, a polysaccha gen, O. —CO.R. —COR, C-Clo alkyl, C-Coalkenyl, ride, or an oligonucleotide. aryl, heteroaryl, C-C cycloalkyl and C-Cls heterocy 50 As used herein, “O-bonded’ refers to a lone pair of cloalkyl; electrons from oxygen being coordinated to M, where M is wherein R7 and Rare independently selected from C-Co Pt, Pd or Ni. Where “O-bonded” modifies “alkyl or “aryl.” alkyl, C-C alkenyl, aryl, heteroaryl, C-C cycloalkyl the M-coordinating oxygen forms a bond to the respective and C-Cls heterocycloalkyl, and alkyl or aryl, wherein M is Pt, Pd or Ni. An exemplary wherein R is selected from hydrogen, hydroxyl, halogen, 55 “O-bonded alkyl moiety is a methoxy radical. An exem amine, thiol, ether, C-C alkyl, C-C alkenyl, aryl, het plary “O-bonded aryl moiety is a phenol radical. Where eroaryl, C-C cycloalkyl and C-Cls heterocycloalkyl. “O-bonded modifies a heteroatom-containing substituent, In the cited exemplary embodiments, the alkyl- and such as "heterocycloalkyl or "heteroaryl, the M-coordi cycloalkyl-containing Substituents can be branched or nating oxygen forms part of the ring system of the respective unbranched, substituted or unsubstituted, saturated or 60 heterocycloalkyl or heteroaryl, wherein Mis Pt, Pd or Ni. An unsaturated, including combinations of different extents of exemplary “O-bonded heterocycloalkyl is a tetrahydropy Substitution, branching and Saturation. Likewise, the alk ran radical. An exemplary “O-bonded heteroaryl' is a furan enyl-containing Substituents can be branched or unbranched, radical. As used herein, “O-bonded ligand’ refers to an substituted or unsubstituted, different extents of unsatura M-coordinating oxygen forming a bond to a ligand, wherein tion, including combinations of different extents of substi 65 M is Pt, Pd or Ni and ligand includes an amino acid, a tution, branching and Saturation. Furthermore, the cited aryl peptide, a protein, a glycan, a peptidoglycan, a polysaccha substituents include substituted and unsubstituted aryl moi ride, or an oligonucleotide. US 9,499,574 B2 13 14 As used herein, "P-bonded refers to a lone pair of acids include, but are not limited to, amino acids comprising electrons from phosphorous being coordinated to M, where photoactivatable cross-linkers, metal binding amino acids, M is Pt, Pd or Ni. Where “P-bonded” modifies “phosphine.” spin-labeled amino acids, fluorescent amino acids, metal the M-coordinating phosphorous (P) is the phosphorous in containing amino acids, amino acids with novel functional the formula P(R)-(R), where x is 0, 1, 2 or 3 and wherein 5 groups, amino acids that covalently or noncovalently inter M is Pt, Pd or Ni. act with other molecules, photocaged and/or photoisomer An "amino acid refers to any monomer unit that can be izable amino acids, radioactive amino acids, amino acids incorporated into a peptide, polypeptide, or protein. As used comprising biotin or a biotin analog, glycosylated amino acids, other carbohydrate modified amino acids, amino acids herein, the term “amino acid' includes the following twenty comprising polyethylene glycol or polyether, heavy atom natural or genetically encoded alpha-amino acids: alanine 10 Substituted amino acids, chemically cleavable and/or pho (Ala or A), arginine (Arg or R), asparagine (ASn or N), tocleavable amino acids, carbon-linked Sugar-containing aspartic acid (Asp or D), cysteine (CyS or C), glutamine (Glin amino acids, redox-active amino acids, amino thioacid con or Q), glutamic acid (Glu or E), glycine (Gly or G), histidine taining amino acids, and amino acids comprising one or (His or H), isoleucine (Ile or I), leucine (Leu or L), lysine more toxic moieties. (Lys or K), methionine (Met or M), phenylalanine (Phe or 15 The term "peptide' refers to a synthetic or naturally F), proline (Pro or P), serine (Ser or S), threonine (Thr or T), occurring polymer of amino acids having two or more amino tryptophan (Trp or W), tyrosine (Tyr or Y), and valine (Val acid moieties joined together through amide bonds. Example or V). In cases where “X” residues are undefined, these of peptides include glutathione, Substance P, calcitonin, should be defined as “any amino acid.” The structures of Enkephalin, and B-type Natrinuretic Peptide. The term these twenty natural amino acids are shown in, e.g., Stryer “protein’ refers to a synthetic or naturally occurring polymer et al., Biochemistry, 5. Sup.th ed., Freeman and Company that is typically encoded by a gene. Naturally occurring (2002), which is incorporated by reference. Additional peptides are typically derived from proteins by post-trans amino acids, such as selenocysteine and pyrrolysine, can lational processing. For the purposes of this disclosure, also be genetically coded for (Stadtman (1996) “Selenocys peptides and proteins together form the group termed “poly teine.” Annu Rev Biochem. 65:83-100 and Ibba et al. (2002) 25 peptides,” which encompasses all polymers composed of “Genetic code: introducing pyrrolysine. Curr Biol. 12(13): only amino acids. R464-R466, which are both incorporated by reference). The An oligonucleotide, as used herein, refers to a polynucle term "amino acid also includes unnatural amino acids, otide having two or more nucleobases connected together modified amino acids (e.g., having modified side chains through phosphoribosyl bonds, phosphodioxyribosyl bonds, and/or backbones), and amino acid analogs. See, e.g., Zhang 30 peptidyldeoxyribosyl bounds, peptidyldeoxyribosyl bonds, et al. (2004) “Selective incorporation of 5-hydroxytrypto phosphorothiolate containing linkages, among others. Oli phan into proteins in mammalian cells.” Proc. Natl. Acad. gonucleotide includes RNA polymers, DNA polymers, Sci. U.S.A. 101(24):8882-8887, Anderson et al. (2004) “An RNA-DNA mixed polymers, peptide nucleic acid polymers, expanded genetic code with a functional quadruplet codon’ locked nucleic acids polymers and mixtures of the foregoing Proc. Natl. Acad. Sci. U.S.A. 101 (20):7566-7571, Ikeda et 35 polymers. Oligonucleotides can include modified bases, al. (2003) “Synthesis of a novel histidine analogue and its modified intemucleotidyl groups (for example, modified efficient incorporation into a protein in vivo. Protein Eng. ribose or deoxyribose moieties and/or modified phosphate, Des. Sel. 16(9):699-706, Chin et al. (2003) “An Expanded phosphorothiolate, or peptide moieties) and/or modified 5' Eukaryotic Genetic Code.” Science 301 (5635):964-967, and 3' terminal groups (for example, a blocking 3'-C3 Spacer James et al. (2001) “Kinetic characterization of ribonuclease 40 group). S mutants containing photoisomerizable phenylaZophenyl Exemplary embodiments of formulas (I)-(VII) include alanine residues.” Protein Eng. Des. Sel. 14(12):983-991, compounds (1)–(19) as illustrated below. Kohrer et al. (2001) “Import of amber and ochre suppressor tRNAS into mammalian cells: A general approach to site specific insertion of amino acid analogues into proteins.” 45 (1) Proc. Natl. Acad. Sci. U.S.A. 98(25): 14310-14315, Bacher et al. (2001) “Selection and Characterization of Escherichia coli Variants Capable of Growth on an Otherwise Toxic Tryptophan Analogue.” J. Bacteriol. 183(18):5414-5425, Hamano-Takaku et al. (2000) “A Mutant Escherichia coli 50 Tyrosyl-tRNA Synthetase Utilizes the Unnatural Amino Acid AZatyrosine More Efficiently than Tyrosine.” J. Biol. (2) Chem. 275(51):40324-40328, and Budisa et al. (2001) “Pro teins with beta}-(thienopyrrolypalanines as alternative chromophores and pharmaceutically active amino acids.” 55 Protein Sci. 10(7): 1281-1292, which are each incorporated C-CH2CH3, by reference. To further illustrate, an amino acid is typically an organic acid that includes a Substituted or unsubstituted amino (3) group, a Substituted or unsubstituted carboxy group, and one 60 or more side chains or groups, or analogs of any of these groups. Exemplary side chains include, e.g., thiol, seleno, Sulfonyl, alkyl, aryl, acyl, keto, azido, hydroxyl, hydrazine, cyano, halo, hydrazide, alkenyl, alkynl, ether, borate, boro nate, phospho, phosphono, phosphine, heterocyclic, enone, 65 imine, aldehyde, ester, thioacid, hydroxylamine, or any combination of these groups. Other representative amino

US 9,499,574 B2 17 18 -continued The preparation of compounds of formula (VII) include (19) the method according to scheme (3).

(scheme 3)

H H N / X N / X 2 O 4. -Y-yw 10 M - S - R-C C-R2, x^ Y R2CN/H2O V - - ?/ O-Q-O C2+ /V HO OH

The synthesis of the disclosed compounds can be accom 15 wherein “RCN represents a mixture of RCN and RCN plished with minor modifications of the following synthetic and "C" represents either two monovalent cations or a procedures. One procedure pertains to an unexpected syn single divalent cation. thesis outcome that uses relatively prolonged, high tempera The preparation of compounds of formulas (I)-(VII), ture conditions than normally done to prepare liganded M-Q wherein the X substituent is replaced with a different X coordination complexes, wherein M is Pt, Pd or Ni and Q is Substituent can be done in several ways. In one embodiment, AS, Sb or Bi. Typically, such liganded M-Q coordination wherein the X substituent contains both an moiety having complexes are synthesized under mild temperature condi N-bonded and S-bonded potential, the alternative X sub tions. Such as ambient or slightly supra-ambient temperature stituent can be obtained by thermal isomerization, as conditions (~15° C. to ~37° C.) to preserve the oxidation depicted in scheme (4) for S-bonded SCN conversion to state of the M of the M-Q coordination complexes. Unusu N-bonded NCS. ally, the some of the M-Q coordination complexes could 25 only be achieved following incubation at 90° C. for ~72 hours. For example, compound (1) can be synthesized by (scheme 4) heating cisplatin with ASOs in an acetonitrile-water mixture (9:1, v/v) at 90° C. for three days. However, the yield of H N H H s H compound (1) increases from 23% to 75% when the starting 30 Y-M-N. N -M-N. material is K-PtCla. I A. y I A. y Alternative, milder synthetic procedures were discovered R- V -R R- V C-R, that enabled other species of the disclosed compounds to be obtained. Exemplary syntheses of the disclosed compounds 8 : V 6 8 : V 6 are provided herein. For example, the preparation of com 35 HO OH HO OH pounds of formula (II) include the method according to wherein “RCN represents a mixture of RCN and RCN scheme (1). and "C" represents either two monovalent cations or a single divalent cation. scheme 1 40 In another embodiment, the replacement of X substituents can be achieved by an exchange reaction, wherein one X H.N / H Substituent, such as a halogen can be replaced by an non X x2 N-M-N. halogen substituent, such as an X substituent having an N / Q(OR34)3 ? M N-bonded group or an S-bonded group according to the /\ RCNCN/H2O R- V -R. 45 schemes (5) and (6). X X of-6 C2+2 R3O MOR wherein "Q(ORs),” represents a mixture of Q(OR), and (scheme 5) Q(OR), and "RCN represents a mixture of RCN and 50 RCN and "C" represents either two monovalent cations HN CI /H 21 N-M-N. or a single divalent cation. A. M s The preparation of compounds of formula (VI) include R-C C-R2 --> the method according to scheme (2). Y, Q { 55 /V HO OH (scheme 2) 21 F. H X X 2 N - MY - N^ s 60 H H N / O A. M N / M - S - R-C C-R2, -M-N. 4 y C2+ /V R'-C C-R HO OH Y, Q { /V wherein “RCN' represents a mixture of RCN and RCN 65 HO OH and "C" represents either two monovalent cations or a single divalent cation. US 9,499,574 B2 19 20 isomers signals are based on multidimensional Pt and 'N scheme 6 NMR spectroscopy on sample of compound (5) which is synthesized using thiocyanate enriched in C and 'N at 21 99% (FIG. 2). C Initial formation of compound (5) with S-bound thiocya HN / H Sa N nate can be kinetically or thermodynamically controlled, but -M-N. both isomers (that is, compounds (5) and (6)) are sufficiently R-c' c-R2 - S - stable in DDMSO solution to be observed using NMR V spectroscopy. The N-isomer of compound (5) is enthalpi O-Q-O 10 cally favored in solution by 15.7 kJmol'. Interestingly, only M HO OH the S-linked complex of compound (5) could be isolated in the solid state, which may be the result of both rapid equilibration and a lower solubility for the S-isomer. Biological Activity of Disclosed Compounds as Anticancer 15 Agents Compounds (1) and (2) demonstrate significant anticancer activity in a panel of human cancer cell lines (FIG. 3 and Example 22) and also overcomes one of the most significant limitations of platinum drugs, namely tumor-based drug 20 resistance mechanisms. The ovarian cisplatin resistant A2780 cancer cell line is of special interest since it encompasses all of the known major mechanisms of resis tance to cisplatin (reduced uptake, increased level of gluta For example variations on formula (VII) (exemplary thione, increased DNA repair, and tolerance to Pt(II)-in species: compound (1)) are accessible by varying Substituent 25 duced lesions). The results show that compound (1) on the nitrile. For instance, compound (2) can be obtained exhibited more than twice the cytotoxicity of cisplatin from the reaction of K-PtCl with ASO in the presence of against the cisplatin resistant cell line A2780 (ICso propionitrile. Conditions for synthesis of compound (2) 21.4+1.8 uM versus 47.3+2.1 uM) (See FIG.3 and Example were different from compound (1) because of the different 22, Table 8). The ability of compound (1) to circumvent miscibility of propionitrile in water (1:9, V/v). Compound 30 cisplatin-acquired resistance was determined from the resis (2) is obtained at mom temperature after 4 days, whereas tance factor (RF), and an RF value of <2 denotes no compound (1) is obtained at elevated temperatures. cross-resistance. In the case of ovarian A2780 and A2780 In compounds (1) and (2), the Pt(11) center adopts a cell lines all approved platinum drugs have RFs between 6.1 square planar geometry, with arsenic, chloride, and two and 16.0. The RF of 1.1 for compound (1) indicates that it nitrogen donors in a trans configuration. The nitrogen donors 35 is far more effective at killing this cisplatin resistant cancer are derived from acetamide (propanamide) formed via Pt assisted acetonitrile (propionitrile) hydrolysis in situ. The cell line and may be able to bypass drug resistance mecha Pt N bond lengths in 1a (Pt1-N1=2.000(3) A and Pt1 nism(s) that lower cisplatin cytotoxicity. N2=2004(3) A are consistent with the Pt N bond lengths Compound (1) has showed better cytotoxic activity than obtained in other Pt(II) complexes with the deprotonated either cisplatin or ASO in colon HCT-116 (ICs-1.6+0.4 form of acetamide, Pt. N=2.004(11) and Pt N=2.000(3) 40 uM vs. 5.5+1.3 uM and 9.4+0.9 uM) and glioblastoma U-87 A. Similar bond lengths are obtained in compound (1b) (ICso-0.37+0.11 uM vs. 9.6+0.8 uM and 1.6+2.9 uM) crystal form b and compound (2) (see Example 21). The cancer cell lines (FIG. 3 and Example 22, Table 8). Addi N1-C1 and O1-C1 bond lengths (both 1.302(4) A) and tionally, compound (1) showed twice the cytotoxicity of N2-C3 and O2-C3 (1.289(6) and 1.297(6) A) in 1a are cisplatin against MDA-MB-231-mCherry cells indicative of a high degree of delocalization present in the 45 (ICso-9.5+0.1 uM and 22.3+2.8 LM), as well as improved chelate rings formed by bridging N, O acetamido ligands cytotoxicity compared with ASO in RPMI 8226 multiple (Example 21). myeloma cells (ICs-4.5+1.0 uM vs. 7.1+0.2 uM, Table 8). The M-Q core is also stable to ligand substitution reac Trans-platinum compounds in comparison with cis-com tions. In general, for example, hydrolysis of Pt—Cl bonds is pounds display different patterns of ligand Substitution, slow (t=2 hat 37°C. and 4 mM Cl) and rapid substitution 50 which contributes to the potency of trans-platinum com usually requires addition of reagents such as AgNO. The pounds in cisplatin-resistant cell lines. The distinct biologi substitution of the Cl ligand in compound (1) with SCNT in cal activity of compound (1) and related compounds in vitro water occurs immediately at room temperature to provide may be the result of the strong trans effect of the As(OH), compound (5) and is likely driven by the trans effect of the moiety combined with the trans stereochemistry of the arsenic moiety. N-atoms at the platinum center. Solution NMR and X-ray crystallography confirm that the 55 The present disclosed contemplates conjugates of the Pt. As bond remained intact. Crystals suitable for a single disclosed compounds coupled to at least one ligand. Suitable crystal X-ray analysis were obtained when compound (5) ligands include an amino acid, a peptide, a protein, a glycan, was synthesized in a 1:1 water/methanol mixture, where the a peptidoglycan, a polysaccharide, or an oligonucleotide. SCN ion is bound to Pt(II) through the sulfur atom (FIG. Such ligands form conjugates with the disclosed compounds 1D). NMR spectroscopy reveals facile linkage isomerization 60 by coordinating with M, where M is Pt, Pd or Ni. Coordi of compound (5) in Solution at mom temperature. Specifi nation with M can occur via N-bonded, S-bonded, O-bonded cally, upon dissolving compound (5) in DDMSO solution or P-bonded coordinations. Alternatively, one of R'-R may equilibrium mixture of 64+1.2% of S-isomer (compound be derivatized to enable covalent bond formation with the (5)) and 36%+1.5 of N-isomer (compound (6)) is quickly ligand. Two exemplary ligand conjugate methods are established, i.e., the "H NMR spectrum obtained after 5 min 65 described in detail below, though one skilled in the art would upon dissolution of compound (5) does not change over recognize similar conjugate chemistry is known for coupling time. Assignments of chemical shifts for the S- and N-bound other ligands described herein. US 9,499,574 B2 21 22 Polypeptide Conjugates If present, the free sulfhydryl group of an available The disclosure encompasses peptide conjugates with the cysteine may form mixed disulfide derivatives with other disclosed compounds. For example, cysteine residues can be thiol-containing compounds, such as other disulfide com reacted, forming disulfide bonds that cross-link protein to pounds. Alternatively, cysteine sulfhydryl functionality may one or more ligands of the disclosed compounds. If the serve as a nucleophile to react with a halide-containing protein does not contain cysteine residues, the protein may compound, such as an alkyl halide or a haloacetamide, or be modified with 2-iminothiolane (Traut’s reagent) using the with a maleimide to form a thioether. chemical scheme (7) shown below. Although the chemical reactivity of alcohol hydroxyl groups of threonine, serine, and tyrosine is low in aqueous 10 Solution, these groups may be selectively modified, espe scheme 7 cially if they are reactive groups within enzyme active sites. Certain N-terminal Seine or threonine groups that exist in a NH non-acylated form in proteins may be oxidized with perio date to yield aldehydes, which can be modified with a variety of amine or hydrazine derivatives. Still other alcohol Protein-NH2 S -e- 15 hydroxyl groups can be selectively modified, like the trip eptide sequences of certain peptides wherein serine, threo Traut's reagent nine or tyrosine residues are separated from a histidine Protein-HN residue by a single amino acid (e.g., Ser-X-His, Thr-X-His r-r and Tyr-X-His), by succinimidyl or sulfosuccinimidyl esters NH or by N-succinimidyl-3-(4-hydroxy-5-'Iliodophenyl) propionate (Bolton-Hunter reagent). The alcohol functionality of tyrosine may be selectively The latter embodiment provide a S-bonded ligand that can modified in several ways. As an indirect method, these coordinate to M of the disclosed compounds, wherein M is groups may be subjected to an initial nitration of the ortho Pt, Pd or Ni. Alternatively, the resultant ligand can be used 25 position of its phenol using tetranitromethane, followed by to couple to other moieties of the disclosed compounds. Such reduction of the o-nitrotyrosine with sodium dithionite as suitably activated R'-R groups as such groups exist in the (Na2SO4) to form an o-aminotyrosine. The resultant aro disclosed compounds. Such embodiments may be advanta matic amine of o-aminotyrosine can react with most amine geous in particular contexts where preservation of the pro reactive reagents. In another approach, the phenol group in tein may serve an important secondary function, such as 30 tyrosine residues may be converted to salicylaldehyde ligand-mediated targeting of the disclosed compounds to derivatives, followed by a reaction of the resultant salicyl ligand-specific receptors located on surfaces of specific cell aldehydes with amine or hydrazine derivatives to yield the types such as cancer cells. modified protein Surfaces. One versatile feature of many embodiments of com The carboxylic groups of aspartate or glutamate may be pound-protein conjugates is that the native structure of the 35 coupled to hydrazines or amines in aqueous solution using water-soluble carbodiimides such as 1-ethyl-3-(3-dimethyl protein is not absolutely critical for compound-protein con aminopropyl)-carbodiimide (EDAC). Including N-hydroxy jugate fabrication. As described previously, stable com Succinamide or N-hydroxysulfoSuccinimide in the reaction pound-protein conjugate formation depends only upon the mixture may improve the coupling efficiency of EDAC presence of reactive sulfhydryl groups in the protein for the mediated protein-carboxylic acid conjugations. To reduce purposes of cross-linking to the disclosed compounds. 40 intra- and inter-protein coupling to lysine residues, which is The compound conjugates can be derivatized to vary the a common side reaction, carbodiimide-mediated coupling in vivo pharmacokinetics and biodistribution of compound may be performed in a concentrated protein solution at a low conjugates. Towards this goal, compound conjugates can pH, using a large excess of the nucleophile. include, but are not limited to, polyethylene glycol chains The amine groups of lysine, glutamine, and arginine may (PEG) (to extend the lifetime of the compound conjugates in 45 form amide linkages following coupling to reactive ester the blood pool), membrane receptor ligands (e.g., folate, compounds. Alternatively, these amine groups may serve as hemes, steroids, neurotransmitters, piperidine-based sigma general nucleophiles with compounds having appropriate receptor ligands), bioactive peptides, and even antibody leaving group reactive chemistry, such as alkyl halides or chains. In these examples, the ligand of interest is cova maleimides. lently-attached to the disclosed conjugates through side 50 Photochemical reagents may represent an alternative chains of amino acid residues coordinated to M, wherein M strategy for modifying the Surfaces of polypeptides for is Pt, Pd or Ni, or to one of the R'-R moieties. conjugation to the disclosed compound(s). Particularly use The availability of numerous functionalizable side groups ful for the present disclosure are multi-functional photore in proteins makes it possible to fabricate protein-compound agents having at least one photoreactive functionality and at conjugates. Only three requirements for Surface modifica least one non-photoreactive chemical functionality. tion of side group functionality must be met. First, some of 55 Examples of groups with a photoreactive functionality the side chains of the amino acids that form the protein must include awl azides and benzophenone derivatives. Examples be accessible to solvent to undergo reaction with the modi of groups with a non-photoreactive chemical functionality fication chemistry. Side chains groups that are buried within include Sulthydryls, amines, alcohols, esters, carbonyls, car the interior of the protein or that lie within the protein boxylates, and halides. The photoreagent may be coupled to tertiary structure may not be solvent accessible to the 60 a polypeptide using an irradiation source corresponding to modification chemistry. Second, and related to the first the W of the photoreactive species. The photoreaction requirement, the target functionality of the side chain must may proceed by a radical reaction, wherein the photoreactive not reside in an environment of secondary or tertiary struc species forms a covalent bond with any amino acid residue ture that may sterically hinder the reaction with the modi in the protein. The photoreactive species may form covalent fication chemistry. Third, and most importantly, the func 65 bonds with tyrosine, phenylalanine, or tryptophan of the tionality of the side group must remain chemically reactive polypeptide, should the wavelength of excitation overlap the to permit conjugation with the disclosed compound(s). respective W of the photoreactive species and the W of US 9,499,574 B2 23 24 any of these amino acid residues. Subsequent conjugation to -continued the disclosed compound(s) may be effected using the sec N C C N ondary chemical functionality attached to the photoreagent. Modification of disclosed compound(s) allows for pro Y- NS longed circulation in the case of polyethylene glycol (PEG) attachment, and Surface recognition in the case of folate and 5 antibodies attachment. “Polyethylene glycol includes poly N- - - mers of ethylene glycol, and moieties and compounds con C C taining —(CH2CH2O)— units, preferably with a mass of at Cyanuric Chloride least 150 daltons, preferably at least 3,000 daltons. Polyethylene glycol (PEG) may be covalently attached to 10 H2N-Protein amine or hydroxyl groups of R'-R of the disclosed com Protein-HN N O(CH2CH2O).H pounds. The amino moiety of lysine and glutamine resides can be modified to introduce functionality such as the NS polyethylene glycol (PEG) group. Likewise, the hydroxyl N N moiety of serine, threonine, and tyrosine may be modified to 15 introduce similar functionality as observed with the amino C moiety. The introduction of the PEG group may be done via a coupling reaction with cyanuric chloride, which is reactive Surface modification using folate is an extension of the with both amino and hydroxyl moieties. The reaction PEG surface modification method shown above. The folate scheme (8) with an amine moiety on the cross-linked in this instance may be activated using N-hydroxySuccina polypeptide is shown below. mide in a dicyclohexyl carbodiimide (DCC) coupling reac tion. The activated folate then may be coupled to a PEG scheme 8 moiety containing an amino group functionality to form an 25 amide bond between PEG and folate. This process is depicted below (scheme (9)). The resultant modified PEG H -e-N-NOH -- can be attached to the Surface of the disclosed compound(s) PEG in the same manner as depicted above via a cyanuric chloride coupling reaction.

scheme 9

HN N N

N 2 N N H Q OH O N

O

HO O Folic acid

DCC N-1- O -- H OH N N N Oa - N - O / 2 N NY N N H Q NHS OH O N

H O Folate-NHS O O

-- O HN N-(N- O n1n 1-)-n NH2 NH-PEG-NH O US 9,499,574 B2 25 26 This example represents one embodiment whereby the through reaction with EDC. The monomeric forms of the disclosed compound(s) may be modified to contain any protein containing EDC-activated carboxylic ester on its ligand using a combination of the preceding chemical reac Surface may be purified and Subsequently reacted with a tion schemes. Thus, one may chemically attach PEG to the suitably derivatized R'-R of the disclosed compounds. The desired ligand using known coupling chemistry. The PEG 5 lysine amino moiety of the polypeptide-compound conju ligand derivative is then coupled to cyanuric chloride and gate surface forms a covalent isopeptide bond with a glu the resultant cyanuric-PEG-ligand compound is reacted with tamate residue in target antibody protein. Optionally, the available amines of the polypeptide to yield the desired order of the reactions may be reversed wherein EDC is Surface modifications. Although the preceding reaction initially reacted with carboxylic acid moieties in amino acid schemes may permit mono-substituted PEG derivatives to 10 residues of the cross-linked polypepide-compound conju be attached at each site of modification in the polypeptide, gate, followed by secondary reaction with lysine moieties in the presence of three functionally reactive chlorines in amino acid residues of the antibody protein ligand to form cyanuric chloride may allow for more extensive coupling the compound conjugated with an antibody. In this fashion, with the PEG derivative. The use of cyanuric chloride may the coupling reaction may be used to covalently like the be advantageous in those cases where each Surface modifi 15 disclosed compounds to any protein that contains available cation on polypeptide may bear a di-substituted PEG deriva carboxylic acid or amino moieties. Furthermore, carboxylate tive. Optionally, one may use any bifunctional protein moieties on the protein my be used for Surface modification crosslinking reagent instead of cyanuric chloride and react by reaction with amines, or by esterification. the resultant PEG-ligand compound with any type of nucleo Oligonucleotide Conjugates phile present on the polypepide. The disclosure encompasses oligonucleotide conjugates The conjugation of folate to the disclosed compounds that include the disclosed compounds. For example, oligo allows targeting to folate-binding tumor cells. Ovarian, nucleotides can be prepared that contain phosphorothiolate breast, and human nasopharyngeal tumors all possess a high groups at one or more internucleotidyl phosphate linkages. concentration of folate receptors on their surface. Liposomes The sulfur moiety of the phosphorothiolate can form an modified by folate-PEG conjugates target folate receptor 25 S-bond with M of the disclosed compounds, wherein M is bearing KB tumor cells and exhibit an inhibitory effect on Pt, Pd or Ni. Alternatively, the sulfur moiety can be used to their growth. Specifically, these types of agents are expected conjugate to one of suitably derivatized R'-R groups of the to target induced squamous cell carcinoma with the folate disclosed compounds. Furthermore, modifications can be modified compounds. Similarly, many oral and upper gas introduced into the terminal positions of oligonucleotides trointestinal tract tumors have a high affinity for various 30 during their synthesis, wherein the modifications include hemes (which greatly assists in the use of hematoporphyrins one or more of an amine or a Sulfur. The resultant oligo as photodynamic therapy agents). Polypeptides with surface nucleotides so modified to contain an amine group can be hemes attached to the disclosed compound(s) may also be used for form an N-bond with M of the disclosed com used to target induced squamous cell carcinoma. pounds, wherein M is Pt, Pd or Ni. Alternatively, the amine The modification of the disclosed compounds with immu 35 moiety can be used to conjugate to one of Suitably deriva noglobulins allows targeting of T-cell receptors. This modi tized R'-R groups of the disclosed compounds. For those fication may be carried out using monoclonal antibodies that oligonucleotides that include terminal modifications that are specific for T-cell receptors. These monoclonal antibod introduce a Sulfur group, then conjugates containing the ies may be covalently coupled to the disclosed compounds disclosed compounds coupled to the resulting oligonucle via a dimethylaminopropyl-carbodiimide hydrochloride 40 otides can be prepared as described supra, wherein the sulfur (EDC) coupling reaction according to scheme (10). group can forman S-bond to M of the disclosed compounds,

scheme 10

GE) CHCH-NH-C=N-(CH)-N(CH) -- Glutamate OH EDC

1. CHCH

Glutamate ision usO I -CH2CH3 Glutamate NH-Lysine + ca C

In this coupling reaction, the carboxylic acid moiety of a 65 wherein M is Pt, Pd or Ni, or wherein the sulfur group can glutamate residue in the protein to be attached to one or be used to conjugate to one of suitably derivatized R'-R more of the disclosed compound(s) may be activated groups of the disclosed compounds. US 9,499,574 B2 27 28 Pharmaceutical Compositions magnesium carbonate, etc. Such compositions and dosage The present disclosure contemplates pharmaceutical com forms will contain a prophylactically or therapeutically positions of preparations of the disclosed compound(s) for effective amount of a preparation of the described com administration to mammals to treat cancer and related con pounds having anti-cancer effect, together with a suitable ditions. In a preferred embodiment, a composition for amount of carrier so as to provide the form for proper administration is a pharmaceutical composition, preferably administration to the patient. in a single unit dosage form. Pharmaceutical compositions The formulation should suit the mode of administration. and single unit dosage forms can comprise a prophylacticly In a preferred embodiment, the pharmaceutical composi or therapeutically effective amount of one or more prophy tions and single unit dosage forms are sterile and prepared lactic or therapeutic agents, and a typically one or more 10 in a form suitable for administration to a subject, preferably pharmaceutically acceptable carriers or excipients or an animal Subject, more preferably a mammalian Subject, diluents. and most preferably a human Subject. Besides humans, The term “pharmaceutically acceptable” means approved preferred animal Subjects include horses, birds, cats, dogs, by a regulatory agency of the Federal or a state government rats, hamsters, mice, guinea pigs, cows, and pigs. (for example, the U.S. Food and Drug Administration) or 15 A pharmaceutical composition of the invention is formu listed in the U.S. Pharmacopeia or other generally recog lated to be compatible with its intended mute of adminis nized pharmacopeia for use in animals, and more particu tration. Examples of mutes of administration include, but are larly in humans. not limited to, parenteral, e.g., intravenous, intradermal, The term “carrier refers to a diluent, adjuvant, excipient, Subcutaneous, intramuscular, Subcutaneous, oral, buccal, or vehicle with which the therapeutic is administered. Such Sublingual, inhalation, intranasal, transdermal, topical, pharmaceutical carriers can be sterile liquids, such as water transmucosal, intra-tumoral, intra-synovial and rectal and oils, including those of petroleum, animal, vegetable or administration. In a specific embodiment, the composition is synthetic origin, such as peanut oil, Soybean oil, mineral oil, formulated in accordance with routine procedures as a sesame oil and the like. Water is a preferred carrier when the pharmaceutical composition adapted for intravenous, Sub pharmaceutical composition is administered intravenously. 25 cutaneous, intramuscular, oral, intranasal or topical admin Saline solutions and aqueous dextrose and glycerol solutions istration to human beings. In an embodiment, a pharmaceu can also be employed as liquid carriers, particularly for tical composition is formulated in accordance with routine injectable solutions. Examples of Suitable pharmaceutical procedures for oral administration to human beings. Typi carriers are described in “Remington's Pharmaceutical Sci cally, compositions for oral administration are solid dosage ences” by E. W. Martin. Pharmaceutical compositions can, 30 forms or solutions in Sterile isotonic aqueous buffer. but need not, comprise one or more active or inactive Examples of dosage forms include, but are not limited to: ingredients that are not necessarily considered pharmaceu tablets; caplets; capsules, such as soft elastic gelatin cap tically acceptable to current practitioners in the art. Sules or hard capsules; dropping pills; cachets; troches; A pharmaceutical composition can be administered by lozenges; dispersions; Suppositories; ointments; cataplasms any mute according to the judgment of those of skill in the 35 (poultices); pastes; powders; dressings; creams; plasters; art, including but not limited to orally, intravenously, intra Solutions; patches; aerosols (for example, nasal sprays or gastrically, intraduodenally, intraperitoneally or intracere inhalers); gels; liquid dosage forms suitable for oral or broventricularly. mucosal administration to a patient, including Suspensions Typical pharmaceutical compositions and dosage forms (for example, aqueous or non-aqueous liquid Suspensions, comprise one or more excipients. Suitable excipients are 40 oil-in-water emulsions, or a water-in-oil liquid emulsions), well-known to those skilled in the art of pharmacy, and Solutions, and elixirs, liquid dosage forms suitable for par non-limiting examples of Suitable excipients include starch, enteral or intravenous administration to a patient; and sterile glucose, lactose. Sucrose, gelatin, malt, rice, flour, chalk, Solids (for example, crystalline or amorphous Solids or silica gel, Sodium Stearate, glycerol monostearate, talc, granular forms) that can be reconstituted to provide liquid Sodium chloride, dried skim milk, glycerol, propylene, gly 45 dosage forms suitable for parenteral or intravenous admin col, water, ethanol and the like. Whether a particular excipi istration to a patient. ent is suitable for incorporation into a pharmaceutical com The composition, shape, and type of dosage forms of a position or dosage form depends on a variety of factors well preparation of the described compounds will typically vary known in the art including, but not limited to, the way in depending on their use. For example, a dosage form used in which the dosage form will be administered to a patient and 50 the acute treatment of a cancer or related disorder may the specific active ingredients in the dosage form. The contain larger amounts of one or more of a preparation of the composition or single unit dosage form, if desired, can also disclosed compounds than a dosage form used in the chronic contain minor amounts of wetting or emulsifying agents, or treatment of the same disease. Also, the therapeutically pH buffering agents. effective dosage form may vary among different types of The disclosure further encompasses administration of 55 diseases or disorders. Similarly, a parenteral dosage form pharmaceutical compositions and single unit dosage forms may contain Smaller amounts of one or more of the active that comprise one or more compounds that reduce the rate by than an oral dosage form used to treat the same disease or which an active ingredient will decompose. Such com disorder. These and other ways in which specific dosage pounds, which are referred to herein as “stabilizers.” forms encompassed by this invention will vary from one include, but are not limited to, antioxidants such as ascorbic 60 another will be readily apparent to those skilled in the art. acid, pH buffers, or salt buffers. See, e.g., Remington's Pharmaceutical Sciences, 18th ed., The pharmaceutical compositions and single unit dosage Mack Publishing, Easton Pa. (1990). forms can take the form of solutions, Suspensions, emulsion, Generally, the ingredients of compositions comprising a tablets, pills, capsules, powders, Sustained-release formula preparation of the disclosed compounds are Supplied either tions and the like. Oral formulation can include standard 65 separately or mixed together in unit dosage form, for carriers such as pharmaceutical grades of mannitol, lactose, example, as a dry lyophilized powder or water free concen starch, magnesium Stearate, sodium saccharine, cellulose, trate in a hermetically sealed container Such as an ampoule US 9,499,574 B2 29 30 or Sachette indicating the quantity of active agent. Where the Disintegrants are used in the compositions of the inven composition is to be administered by infusion, it can be tion to provide tablets that disintegrate when exposed to an dispensed with an infusion bottle containing sterile pharma aqueous environment. Tablets that contain too much disin ceutical grade water or saline. Where the composition is tegrant may disintegrate in storage, while those that contain administered by injection, an ampoule of sterile water for too little may not disintegrate at a desired rate or under the injection or saline can be provided so that the ingredients desired conditions. Thus, a Sufficient amount of disintegrant may be mixed prior to administration. that is neither too much nor too little to detrimentally alter Pharmaceutical compositions used in the methods of the the release of the active ingredients should be used to form invention that are suitable for oral administration can be solid oral dosage forms of the invention. The amount of presented as discrete dosage forms, such as, but are not 10 disintegrant used varies based upon the type of formulation, limited to, tablets (e.g., chewable tablets), caplets, capsules, and is readily discernible to those of ordinary skill in the art. and liquids (e.g., flavored syrups). Such dosage forms con Typical pharmaceutical compositions comprise from about tain predetermined amounts of active ingredients, and may 0.5 to about 15 weight percent of disintegrant, specifically be prepared by methods of pharmacy well known to those from about 1 to about 5 weight percent of disintegrant. skilled in the art. See generally, Remington’s Pharmaceuti 15 Disintegrants that can be used in pharmaceutical compo cal Sciences, 18th ed., Mack Publishing, Easton Pa. (1990). sitions and dosage forms of the invention include, but are not In certain embodiments, the oral dosage forms are solid limited to, agar-agar, alginic acid, calcium carbonate, micro and prepared under anhydrous conditions with anhydrous crystalline cellulose, croScarmellose sodium, crospovidone, ingredients, as described in detail in the sections above. polacrilin potassium, Sodium starch glycolate, potato or However, the scope of the invention extends beyond anhy tapioca starch, pre-gelatinized starch, other starches, clays, drous, Solid oral dosage forms. As such, further forms are other algins, other celluloses, gums, and mixtures thereof. described herein. Lubricants that can be used in pharmaceutical composi Typical oral dosage forms are prepared by combining the tions and dosage forms of the invention include, but are not active ingredient(s) (that is, a preparation of the he disclosed limited to, calcium Stearate, magnesium Stearate, mineral oil, compound(s)) in an intimate admixture with at least one 25 light mineral oil, glycerin, Sorbitol, mannitol, polyethylene excipient according to conventional pharmaceutical com glycol, other glycols, Stearic acid, sodium lauryl Sulfate, talc, pounding techniques. Excipients can take a wide variety of hydrogenated vegetable oil (e.g., peanut oil, cottonseed oil, forms depending on the form of preparation desired for Sunflower oil, Sesame oil, olive oil, corn oil, and soybean administration. For example, excipients suitable for use in oil), Zinc stearate, ethyl oleate, ethyl laureate, agar, and oral liquid or aerosol dosage forms include, but are not 30 mixtures thereof. Additional lubricants include, for example, limited to, water, glycols, oils, alcohols, flavoring agents, a syloid silica gel (AEROSIL 200, manufactured by W.R. preservatives, and coloring agents. Examples of excipients Grace Co. of Baltimore, Md.), a coagulated aerosol of Suitable for use in Solid oral dosage forms (e.g., powders, synthetic silica (marketed by Degussa Co. of Plano, Tex.), tablets, capsules, and caplets) include, but are not limited to, CAB-O-SIL (a pyrogenic silicon dioxide product sold by starches, Sugars, micro-crystalline cellulose, diluents, granu 35 Cabot Co. of Boston, Mass.), and mixtures thereof. If used lating agents, lubricants, binders, and disintegrating agents. at all, lubricants are typically used in an amount of less than Examples of excipients that can be used in oral dosage about 1 weight percent of the pharmaceutical compositions forms of the invention include, but are not limited to, or dosage forms into which they are incorporated. binders, fillers, disintegrants, and lubricants. Binders Suit The amount of the composition in the methods of the able for use in pharmaceutical compositions and dosage 40 invention which will be effective in the prevention, treat forms include, but are not limited to, corn starch, potato ment, management, or amelioration of a cancer disorder or starch, or other starches, gelatin, natural and synthetic gums one or more symptoms thereof will vary with the nature and Such as acacia, Sodium alginate, alginic acid, otheralginates, severity of the disease or condition, and the mute by which powdered tragacanth, guar gum, cellulose and its derivatives the active ingredient is administered. The frequency and (e.g., ethyl cellulose, cellulose acetate, carboxymethyl cel 45 dosage will also vary according to factors specific for each lulose calcium, Sodium carboxymethyl cellulose), polyvinyl patient depending on the specific therapy (e.g., therapeutic pyrrolidone, methyl cellulose, pre-gelatinized starch, or prophylactic agents) administered, the severity of the hydroxypropyl methyl cellulose, (e.g., Nos. 2208, 2906, disorder, disease, or condition, the mute of administration, as 2910), microcrystalline cellulose, and mixtures thereof. well as age, body, weight, response, and the past medical Examples of fillers suitable for use in the pharmaceutical 50 history of the patient. Effective doses may be extrapolated compositions and dosage forms disclosed herein include, but from dose-response curves derived from in vitro or animal are not limited to, talc, calcium carbonate (e.g., granules or model test systems. powder), microcrystalline cellulose, powdered cellulose, Typical dosage forms for administration in methods of the dextrates, kaolin, mannitol, Silicic acid, Sorbitol, starch, invention comprise a composition of the invention in an pre-gelatinized starch, and mixtures thereof. The binder or 55 amount within the range of from about 0.001 mg to about filler in pharmaceutical compositions of the invention is 500 mg of the disclosed compounds per day, 0.10 mg to 300 typically present in from about 50 to about 99 weight percent mg of the disclosed compounds per day, or 1.0 mg to 200 mg of the pharmaceutical composition or dosage form. of the disclosed compounds per day given as a single Suitable forms of microcrystalline cellulose include, but once-a-day dose or as divided doses throughout the day. are not limited to, the materials sold as AVICEL-PH-101, 60 Particular dosage forms of the invention have incremental AVICEL-PH-103 AVICEL RC-581, AVICEL-PH-105 variations from about 0.001, 0.01, 0.1, 0.2, 0.25, 0.3, 0.5, (available from FMC Corporation, American Viscose Divi 0.75, 1.0, 2.0, 2.5, 3.0, 5.0, 7.5, 10.0, 15.0, 20.0, 25.0, 40.0, sion, Avicel Sales, Marcus Hook, Pa.), and mixtures thereof. 50.0, 60.0, 75.0, 100, 125, 150, 175, 200, 250, 300, 400, and An specific binder is a mixture of microcrystalline cellulose 500 mg of the disclosed compounds, as well as incremental and sodium carboxymethyl cellulose sold as AVICEL 65 dosage variations thereof. RC-581. Suitable anhydrous or low moisture excipients or Exemplary dosage forms of the invention having a liquid additives include AVICEL-PH-103TM and Starch 1500 LM. formulation include 1, 3, 5, 7.5, 10, 15, 20, 50, 75, and 100 US 9,499,574 B2 31 32 ml of a liquid composition of the disclosed compounds The therapeutic index is the dose ratio between therapeu having a concentration ranging from about 0.01 ug/ml to tic effect and toxicity effect. Compounds which exhibit high about 500 mg/ml. The preferred concentrations of such therapeutic indices are preferred. The data obtained from liquid compositions will depend upon the dissolution char these cell culture assays and animal studies can be used in formulating a dosage range that is not toxic for use in acteristics of the medium, which will determine the upper humans. The dosage of the compounds described herein lies limit of pharmaceutically acceptable concentrations of the preferably within a range of circulating concentrations that disclosed compounds in Such compositions. Consequently, include the effective dose with little or no toxicity. The alternative, pharmaceutically acceptable, concentrations of dosage may vary within this range depending upon the the disclosed compounds in liquid compositions that are dosage form employed and the route of administration lower, as well as higher, than that Stated herein are also 10 utilized. The exact formulation, mute of administration and contemplated by the present invention. dosage can be chosen by the individual physician in view of In the case of liquid dosage forms, Suitable concentrations the patient’s condition and the indication to be treated. (See, of the disclosed compounds are Suspended or dissolved in e.g., Finglet al., 1996. In: The Pharmacological Basis of pharmaceutically acceptable carrier media, Such as water, Therapeutics, 9" ed., Chapter 2, p. 29, Elliot M. Ross). 15 The descriptions of exemplary doses are merely alterna saline, and the like. Furthermore, suitable concentrations of tive descriptions that may be used optionally at the discre the disclosed compounds are Suspended or dissolved under tion of the physician and are not intended to conflict or physiologically and physiochemically appropriate condi supersede other descriptions of doses disclosed herein. The tions. pharmacological action of preparations of the disclosed As explained in the disclosure, certain of the disclosed compounds is demonstrated by the disclosed biological compounds differ in their hydrophobicity, thereby rendering examples. them insoluble, slightly soluble, moderately soluble, very EXAMPLES soluble or highly soluble in a particular solvent system, Such as aqueous solvents (for example, water or saline) or solvent Example 1 systems (for example, water/DMSO or saline/DMSO). To 25 the extent that solubility presents a barrier to providing General Methods adequate ADME properties for the delivery system or mute of administration for certain of the disclosed compounds, Cisplatin, KPtCl), DMSO, methanol, acetonitrile, pro particle size reduction of the disclosed compounds to pionitrile and ASOs were purchased from Sigma-Aldrich increase solvent-accessible Surface area and Solvation poten 30 and used without further purification. Media for the cell tial can improve solubility of the disclosed compounds cultures and all other cell Supplements were purchased from having hydrophobic properties. Methods, techniques and Invitrogen. The following spectrometers were used: NMR: instrumentation are known in the art for achieving particle Bruker Avance-III 600 and 500 MHz: chemical shifts (H, size reduction (for example, micronization) can be useful for 'C) were referenced to tetramethylsilane at 0.000 ppm 'H achieving particle size reduction for the disclosed com 35 and 'C if it was present, and to residual solvent signal for samples in which TMS was absent (for DDMSO, 8g250 pounds. ppm and 8–39.5 ppm). The 'Pt (129 MHz) and N (61 Exemplary doses of a composition of the disclosed com MHz) NMR spectra were acquired on a 600 MHz Bruker pounds include microgram or milligram amounts of the Avance III spectrometer equipped with BBO (broadband disclosed compound(s) per kilogram of Subject or sample observe) probe. 'Pt NMR chemical shifts were referenced weight. For a composition used in the invention, the dosage 40 indirectly to TMS in "H NMR spectrum such that N'PtCl administered to a patient can be administered from about in DO would resonate at 0.0 ppm. 'N NMR chemical 0.001 mg/kg to about 10 mg/kg. In certain embodiments, the shifts, both those observed directly in N-detect 1D experi dosage can be from about 0.02 mg/kg to about 4.0 mg/kg of ments and 2D 'H-'N HSQC experiments, were referenced the patient’s body weight, based on weight and purity of a indirectly to TMS in the "H spectra, such that "NHC1 preparation the disclosed compound(s) in the composition. 45 would resonate at 0.0 ppm. The 'Pt NMR spectra were The composition can be administered as a single once-a- acquired with the simple Zg pulse sequence, using an acqui day dose or as divided doses throughout a day. In some sition time of 0.020 sec, and a relaxation delay of 0.1 sec. embodiments, the daily dose is administered twice daily in Spectra acquired for the variable temperature (VT) experi equally divided doses. In other embodiments, the daily dose ments involved setting the temperature to a particular value is administered three times per day. In particular embodi 50 (in 5-degree increments), waiting 15 minutes for equilibra ments, the daily dose is administered three times per day in tion, calibrating the temperature using a neat ethylene glycol equally divided doses. In particular embodiments, the daily temperature calibration standard, inserting the sample of dose is administered four times per day in equally divided interest, equilibrating it for five minutes, then acquiring a doses. The actual dosage can be determined by a practitioner 24-scan "H spectrum employing a 90° pulse and a 30-second of skill in the art according to, for example, the Subjects age, 55 D1 delay for relaxation, ensuring accurate peak integration. body weight, body mass index, or other factors. In certain Data were acquired using Topspin 2.1.4. UV/Vis electron embodiments, administration of a composition in the inven absorbance measurements were performed on a Perkin tion may be repeated daily. In certain embodiments and the Elmer Lambda 650 spectrophotometer. IR: Bruker Tensor 37 administrations may be separated by at least 1 day, 2 days or FT-IR. Elemental analysis was performed at Prevalere, Life 3 days. 60 Sciences, LLC, Whitesboro, N.Y. An effective amount of a composition described herein Example 2 will provide therapeutic benefit without causing substantial toxicity. Toxicity of a composition can be determined by Synthesis of compound (1) (arsenoplatin, Pt(LL standard pharmaceutical procedures in cell cultures or NHC(CH)O)C1As(OH)) experimental animals, for example, by determining the LDso 65 (that is, the dose lethal to 50% of the population) or the Cisplatin (300 mg, 1.00 mmol) or K-PtCl (415 mg. LDoo (that is, the dose lethal to 100% of the population). 1.00 mmol) was added to 125 cm of 9:1 CHCN/HO (v/v). US 9,499,574 B2 33 34 The mixture was stirred at 90° C. Once the platinum 172.6 (CO); 'Pt NMR (129 MHz, DDMSO) 8 -3861 compound was dissolved, 396 mg ASO (2.00 mmol) was ppm (s, 1Pt): UV/Vis (methanol-water-1:1): ||=285 nm. added to the solution, and the reaction mixture was stirred at (E=4217 dm mol' cm):=225 nm (E=15919 dm mol' 90° C. for 72 hours. The resulting mixture was filtered, and cm), shoulder at 265 nm. Solubility: DMSO and methanol. the filtrate was left at room temperature in a glass beaker for 5 approximately 4 weeks until crystals formed. Crystals Suit Example 5 able for a single crystal X-ray analysis were retrieved from solution. The solution was filtered, washed with CHCN: Synthesis of compound (6) (Pt(u-NHC(CH)O) H2O (9:1 mixture), and the obtained crystals were dried in (NCS)As(OH)) a dessicator. Yield 350 mg product (75%; 23% when cis 10 platin is used). Complex is soluble in DMSO, methanol, Compound (5) obtained in Example 4 was dissolved in ethanol, and partially in water. Elemental analysis (% calcd. DMSO solution at room temperature. Isomerization of Com found for C7Hiss ASCINOPt): C, (15.20, 15.24); H. pound (5) occurred. NMR data: "H NMR (600 MHz, D. (3.37, 3.36): N, (8.86, 8.69). NMR: "H NMR (600 MHz, DMSO): 8 2.17 (s, 6H, CH), 8.57 (s. 2H, NH), 9.11 (s. 2H, DDMSO, 25° C) 8: 8.91 (s. 2H-OH), 8.15 (s, 15 OH). 'C NMR (150 MHz, DDMSO) 816.4 (CH), 134.4 2H NH), 2.13 (s, 6H CH). ''C NMR (150 MHz, D. (NCS), 172.7 (CO). 'Pt NMR (129 MHz, DDMSO) & DMSO, 25° C.): 8 172.1 (C-1, C-3), 1714 (C-5, C-7), 22.5 -3724 ppm (s, 1 Pt). 'N NMR (60 MHz, from the "H-'N (C-6, C-8), 16.6 (C-2, C-4). 'N NMR (60 MHz, from the 2D HSQC and N-detect 1D of the sample isotopically 'H-'N HSQC, DDMSO, referenced to 'NHC1, 25° C): labeled with SCN, DDMSO): & 105.8 (N-1, N-2), 91.5 8106.4 (N-1, N-2), 'Pt NMR (129 MHz, DDMSO, 259 ppm (SCN). C.), 6: -3589 (s, 1 Pt) ppm. UV/Vis (methanol-water=1:1): =283 nm (e=4062 dm mol' cm), .247 nm (e=3621 Example 6 dm mol' cm), sh. 260 nm. Synthesis of compound (7) (Pt(u-NHC(CHCH) Example 3 25 O)(SCN)As(OH)) Synthesis of compound (2) (Pt(u-NHC(CHCH.) Compound (2) obtained in Example 3 (96.72 mg, 0.2 O)C1As(OH)) mmol) was dissolved in 4 cm of methanol and added to an equal-molar solution of KSCN (19.44 mg., 0.2 mmol) in 4 K-PtCl(415 mg, 1.00 mmol) was added to 50 cm of 30 cm of water. The reaction mixture was allowed to stir at 50° 9:1 HO/propionitrile (v/v). The mixture was stirred at mom C. for 5 hours. The mixture was filtered and the solution was temperature. Once the platinum compound was dissolved, left standing at mom temperature until crystals appeared. 396 mg ASO, (2.00 mmol) of the reaction mixture was The obtained crystals were removed by filtration. Yield: 37 stirred at room temperature for 96 hours. mg (37%). LC/ESI-MS data were recorded every 24 hours. The color 35 gradually turned yellow and ultimately colorless over the 4 Example 7 day period. The resulting mixture was filtered and left to sit (pH roughly 2) at 25° C., and, within a week, crystals Synthesis of compound (8) (IPt(u-NHC(CHCH.) Suitable for single crystal X-ray analysis appeared. The O) (NCS)As(OH)) solution was filtered again, washed with HO, dried, and 40 weighed (212 mg, 44%). Compound (2) is soluble in metha Compound (7) obtained in Example 6 was dissolved in nol, DMSO, and partially soluble in H.O. DMSO solution at room temperature. Isomerization of com Elemental analysis (% calcd., found for pound (7) occurred and both Compounds (7) and (8) are CHASCINOPt): C, (14.90, 14.80); H. (2.92, 2.60): N, present in the solution. (5.79, 5.82). NMR: "H NMR (500 MHz, DDMSO): 8 45 8.91 (s. 2H-OH), 8.04 (s. 2H NH), 2.47 (q, J=7.6 Hz, Example 8 4H CH), 1.04 (t, J=7.6 Hz, 6H-CH). ''C NMR (125 MHz, DDMSO): & 175.0, 24.1, 11.3 ppm. UV/Vis Synthesis of compound (9) (IPt(u-NHC(CH)O) ((methanol-water=1:1 V/v):=285 nm, a 245 nm. (Py)As(OH)) 50 Example 4 Compound (1) obtained in Example 2 (100 mg, 0.18 mmol) was dissolved in 4 cm of methanol and water Synthesis of compound (5) (Pt(u-NHC(CH)O) mixture, and 14.6 L of pyridine and 22.1 mg of NaClO (SCN)As(OH)) were added. The reaction mixture was allowed to stir at 55 room temperature over night. HRMS spectrum indicated Compound (1) obtained in Example 2 (100 mg, 0.18 presence of M ion at 498 m/z. mmol) was dissolved in 4 cm of methanol and added to an equal-molar solution of KSCN (17.6 mg, 0.180 mmol) in 4 Example 9 cm of water. The reaction mixture was allowed to stir at 50° C. for 5 hours. The mixture was filtered and the solution was 60 Synthesis of compound (10) (Pt(L-NHC(CH)O) left standing at mom temperature until crystals appeared. ClAs(OH)(OCHI) The obtained crystals were removed by filtration. Yield: 60.00 mg (52%). Elemental analysis (% calcd., found for Compound (1) obtained in Example 2 (100 mg, 0.18 PtCHNOSAs): C, (12.56, 12.37); H. (2.11, 1.97); N, mmol) was dissolved in 4 cm of anhydrous methanol. (8.78, 8.56). NMR data: "H NMR (600 MHz, DDMSO): 65 HRMS spectrum indicates molecular ion at m/z. 470 in the 8 2.21 (s, 6H, CH), 7.70 (s. 2H, NH), 9.20 (s. 2H, OH). 'C negative mode consistent with the structure of compound NMR (150 MHz, DDMSO) 817.5 (CH), 117.7 (SCN), (10) was obtained. US 9,499,574 B2 35 36 Example 10 90° C. Once the platinum complex is dissolved, 583 mg SbO, (2.00 mmol) will be added to the solution, and the Synthesis of compound (3) (Pt(u-NHC(CH)O) reaction mixture will be stirred at 90° C. for 72 hours. IAs(OH)) Example 17 KPtII (780 mg, 1.00 mmol) will be dissolved in 125 cm of 9:1 CHCN/HO (v/v). The mixture will be stirred at Synthesis of compound (16) (Pd(LL-NHC(CH)O) 90° C. Once the platinum complex is dissolved, 386 mg CISb(OH)) ASO (2.00 mmol) will be added to the solution, and the KPdCl (326 mg, 1.00 mmol) will be dissolved in 125 reaction mixture will be stirred at 90° C. for 72 hours. cm of 9:1 CHCN/HO (v/v). The mixture will be stirred at 10 90° C. Once the palladium complex is dissolved, 583 mg Example 11 SbO, (2.00 mmol) will be added to the solution, and the Synthesis of compound (4) (Pt(u-NHC(CHCH.) reaction mixture will be stirred at 90° C. for 72 hours. O)IAS(OH)) Example 18 KPtII (780 mg, 1.00 mmol) will be dissolved in 125 15 Synthesis of compound (17) (Ni(u-NHC(CH)O), cm of 1:9 CHCHCN/HO (v/v). Once the platinum com CISb(OH)) plex is dissolved, 396 mg ASO, (2.00 mmol) will be added to the solution, and the reaction mixture will be stirred at KNiCl, (288 mg, 1.00 mmol) will be dissolved in 125 room temperature for four days. cm of 9:1 CHCN/HO (v/v). The mixture will be stirred at 90°C. Once the nickel complex is dissolved, 583 mg SbO Example 12 (2.00 mmol) will be added to the solution, and the reaction mixture will be stirred at 90° C. for 72 hours. Synthesis of compound (11) (Pd(L-NHC(CH)O) Example 19 C1As(OH),) Synthesis of compound (18) (Pt(u-NHC(CH)O) KPdCl (326 mg, 1.00 mmol) will be dissolved in 125 25 (Py)Sb(OH)) cm of 9:1 CHCN/HO (v/v). The mixture will be stirred at 90° C. Once the palladium complex is dissolved, 396 mg Compound (15) obtained in Example 16 (502 mg, 1 ASO (2.00 mmol) will be added to the solution, and the mmol) will be dissolved in 4 cm of methanol and water reaction mixture will be stirred at 90° C. for 72 hours. mixture, and 14.6 L of pyridine and 22.1 mg of NaClO. will be added. The reaction mixture will be allowed to stir 30 Example 13 at mom temperature over night. Example 20 Synthesis of compound (12) (Pd(LL-NHC(CHCH.) Synthesis of compound (19) (Pt(u-NHC(CH)O), O)C1As(OH)) (Thiophenol)Sb(OH)) KPdCl (326 mg, 1.00 mmol) will be dissolved in 125 35 Compound (15) obtained in Example 16 (502 mg, 1 cm of 1:9 CHCHCN/HO (v/v). Once the palladium mmol) will be dissolved in 4 cm of methanol and 102 uL complex is dissolved, 396 mg ASO (2.00 mmol) will be of thiophenol (1 mmol) will be added. The reaction mixture added to the solution, and the reaction mixture will be stirred at mom temperature for four days. will be allowed to stir at room temperature over night. 40 Example 21 Example 14 Crystallographic Structure Determination and Synthesis of compound (13) (Ni(L-NHC(CH)O) Refinement Details ClAs(OH)) Colorless crystals of compounds (1), (2) and (5) were mounted using oil (Infineum V8512) on a glass fiber. All KNiCl, (288 mg, 1.00 mmol) will be dissolved in 125 45 measurements were made on a Broker APEX-II CCD area cm of 9:1 CHCN/HO (v/v). The mixture will be stirred at detector with graphite monochromated MoK\C. radiation. 90°C. Once the nickel complex is dissolved, 396 mg ASO, The data were collected at a temperature of 100(2) K (1a, 2. (2.00 mmol) will be added to the solution, and the reaction and 3) and 111(2) K (1b), and integrated and corrected for mixture will be stirred at 90° C. for 72 hours. decay and Lp effects using Broker APEX II software. Final 50 unit cell parameters were obtained through a refinement of Example 15 all observed reflections during data integration. A face indexed absorption correction was performed via)(PREP. Synthesis of compound (14) (Ni(L-NHC(CHCH.) The structures were solved and refined using the SHELXTL O)C1As(OH)) suite of software. In the structure of compound (1) (crystal KNiCl, (288 mg, 1.00 mmol) will be dissolved in 125 55 form la) the non-hydrogen atoms were refined anisotropi cm of 1:9 CHCHCN/HO (v/v). The mixture will be cally. There is an acetamide and water disordered over the stirred at 90° C. Once the nickel complex is dissolved, 396 inversion center. The hydrogen atoms on the water were not mg ASO, (2.00 mmol) will be added to the solution, and the found in the difference map. The C8 atom (in 1a) and Ni (in reaction mixture will be stirred at room temperature for four 3) were restrained with Uij components approximate to days. isotropic behavior. Hydrogen atoms were included in ideal 60 ized positions, but not refined. In the structure of 1b hydro Example 16 gen atoms on the oxygen and nitrogen atoms were refined isotropically. Neutral atom scattering factors, the values for Synthesis of compound (15) (Pt(LL-NHC(CH)O) Df and Df", and the values for the mass attenuation coef CISb(OH),) ficients were taken from the usual tabulation. Anomalous 65 dispersion effects were included in F. Explanations for B KPtCl(415 mg, 1.00 mmol) will be dissolved in 125 alerts generated from CheckCifare discussed in the refine cm of 9:1 CHCN/HO (v/v). The mixture will be stirred at special details of the corresponding cif files.

US 9,499,574 B2 39 40 TABLE 4-continued Hydrogen bonds for 1b (A and deg.l. D. H. A ARU) d(D–H) d(H A) d(D A)

Crystal data for compound (2). CHASCINOPt, TABLE 7-continued Mr=483.65, Mo KO. radiation, wavelength 0.71073, T 100 (2) K colorless plate, 0.326x0.271 x0.02 mm, orthorhombic, Selected bonds (A) and angles (o) for compound (3). space group Pbca, a=14.1727(5) A, b=9.6476 (3) A, 15 Pt1 As1 2.2987 (9) N2 Pt1 S1 88.6(2) c=17.2048 (6) A, V=2352.46 (14) A, Z=8, dcaica =2.731 Pt1 S1 2.352(2) As1 Pt1 S1 176.70(6) gcm, u=14.944 mm, F(000)=1792, 43,536 reflections, As1 O3 1.720(6) O3 As1 O4 106.6(3) 3424 unique, R=0.0692, R=0.0243 D2O (I), As1 O4 1.727(6) O3 As1 O1 88.0(3) wR=0.0627 (all data), GOF 1094. Representative crystal- A. 8. 2.8 O4 As1 O1 89.4(3) S O3 As1 O2 88.0(3) lographic data for 2 is presented in Tables 5 and 6. 2O S1 C5 1.675 (9) O4 As1 O2 88.5(3) O1 C1 1.316(10) O1 As1 O2 174.8(2) TABLE 5 O2 C3 1.284 (10) O3 As1 Pt1 125.6(2) N1 C1 1.282 (12) O4 As1 Pt1 127.8(2) Selected bonds (A) and angles (deg.) for compound (2). N2 C3 1.303(10) O1 As1 Pt1 93.65 (18) N3 C5 1.162(12) O2 As1 Pt1 91.43 (16) Pt(1) N(1) 1997(3) O(1) C(1) 1.301(4) 25 N1 Pt1 N2 173.9(3) C5 S1 Pt1 104.6(3) Pt(1) N(2) 1997(3) N(1)-C(1) 1.299(4) N1 Pt1 As1 85.8(2) C3 N2 Pt 1 120.4(6) Pt(1)—AS(1) 2.2687 (4) N(1)-H(1) O.88OO Pt(1) Cl(1) 2.3361 (9) N(2) C(4) 1.305(5) As(1)–O(4) 1.724(2) O(3)—As(1)–O(1) 87.15(12) As(1)–O(3) 1.742(3) O(4)–As(1)–O(2) 89.11(12) Example 22 As(1)–O(1) 1.955(3) O(3)-As(1) O(2) 89.99(12) 30 As(1)–O(2) 1976(3) As(1) Pt(1) Cl(1) 177.32(3) As(1)—O(2) 1976(3) O(4)-As(1) O(3) 105.48(13) Cell Culture Conditions and In Vitro Cytotoxicity O(2)—C(4) 1.304(4) O(4)–As(1)–O(1) 85.55(11) Assay N(1)-Pt(1) N(2) 173.59(13) O(1)–As(1) O(2) 173.05 (11) N(1)-Pt(1)—AS(1) 86.22(9) O(4)-As(1) Pt(1) 129.78(10) A. Cell Culture Conditions. N(2)—Pt(1)—AS(1) 87.42(9) O(3)-As(1) Pt(1) 124.67(9) N(1)-Pt(1) Cl(1) 91.71 (9) O(1)–As(1) Pt(1) 93.75(7) The MDA-MB-231-mCherry breast cancer cells were N(2) Pt(1) Cl(1) 94.68(9) O(2)-As(1) Pt(1) 93.09(7) cultured in Dulbecco's modified Eagle's medium (DMEM) C(1) N(1)- Pt(1) 122.0(3) and supplemented with 5% heat-inactivated fetal bovine serum (FBS), 50 units/ml penicillin, 50 g/ml streptomycin, 2 mM L-glutamine, and 1 ug/ml blasticidin S (Sigma). The TABLE 6 Hydrogen bonds for compound (2) [A and degl D. H. A ARU) d(D–H) d(H A) d(D A)

Crystal data for compound (3). CHASNOPtS, A2780 and A2780 ovarian cancer cell lines and the Mr 478.23, Mo KC radiation, wavelength 0.71073, T multiple myeloma RPMI 8226 cell line were cultured in 100(2) K, colorless needle, 0.37x0.08x0.03 mm, orthorhom RPMI 1640 medium supplemented with 10% FBS, 50 bic, space group Pccn, a=17.1342 (5) A, b=18.7310 (5) A, 55 units/ml penicillin, 50 ug/ml streptomycin, 2 mM L-gluta c=6.8347 (2) A, v=2193.53 (11) A. Z=8, d2.896 mine. The U-87 and HTC-116 cancer cell lines were cul gcm, u=15.976 mm, F(000)=1760, 35,638 reflections, tured without antibiotics. Cells were grown at 37° C. in a 3160 unique, R=0.0890, R=0.0460 D2O(I), humidified atmosphere of 5% of CO. wR=0.1117 (all data), GOF 1.046. Representative crystal B. In Vitro Cytotoxicity Assay for MDA-MB-231mCherry, lographic data for 3 is presented in Table 7. 60 A270 and A2780CP Cancer Cell Lines. The cytotoxicities of compound (1), cisplatin, and As2O3 TABLE 7 were assessed by MTS assay using the CelTiter 96 Aqueous MTS (Promega). The 100 uL aliquots of cell suspension Selected bonds (A) and angles (o) for compound (3). (10x10 cells/ml) were plated in 96-well tissue culture Pt1 N1 2.036(7) N2 Pt1 As1 88.19(19) 65 plates in the incubator overnight at 37° C. in a humidified Pt1 N2 2.039(7) N1 Pt1 S1974(2) atmosphere of 5% of CO. The serial dilutions of compound (1), cisplatin, and ASOs in appropriate media were trans US 9,499,574 B2 41 42 ferred to the cells. The MTS solution (20 uL) was added to be inclusive and mean that there may be additional after 72 hours and the absorbance was measured at 495 nm. elements other than the listed elements. The word 'or' 4 hours later. Sigmoidal dose response curves were plotted means any one member of a particular list and also includes using the GraphPad Prism software. The ICs values were any combination of members of that list, unless otherwise obtained on at least three independent experiments. In vitro specified. cytotoxicity assay for U-87 and HTC-116 cancer cell line. The modal verb “may refers to the preferred use or For the experiment, cells were cultured in RPMI 1640 selection of one or more options or choices among several medium with 5% FBS and 2 mM L-glutamine. The drug was described embodiments or features contained within the added in similar media Supplemented with 50 ug/mL gen same. Where no options or choices are disclosed regarding tamicin. HCT-116 and U-87 cancer cell were plated in 96 10 a particular embodiment or feature contained in the same, well plates. The cells were plated in four replicates. The the modal verb “may refers to an affirmative act regarding densities of the cells were 5000 and 10000 per well in 90 uL how to make or use an aspect of a described embodiment or for HTC-116 and U-87, respectively. Ten microliters (10 uL) feature contained in the same, or a definitive decision to use of the final drug volume have added per well. The cells were a specific skill regarding a described embodiment or feature treated with 100, 10, 1, 0.1 0.01, 0.001, 0.0001, 0.00001 and 15 contained in the same. In this latter context, the model verb 0.000001 uM of compound (1), cisplatin, and ASO. After “may has the same meaning and connotation as the auxil 48 hours post drug treatment all plates were allowed to iary verb “can.” equilibrate for 30 minutes at mom temperature from incu The term “about is used herein to mean approximately, bator. Addition of 100 uL of Cell Titer Glow (Promega, roughly, around, or in the region of. When the term “about Fitchburg, Wis.) reagent was added to the wells. The plates is used in conjunction with a numerical range, it modifies were shaken for 2 minutes and allowed to sit for activation that range by extending the boundaries above and below the prior to reading for 10 minutes. Plates were read on Biotek numerical values set forth. Preferably, the term “about is Synergy H1 (Biotek, Wenooski, Vt.) reader capable of used herein to modify a numerical value above and below luminescence detection. the stated value by a variance of 20 percent up or down 25 (higher or lower). TABLE 8 The terms “disclosed compound,” “disclosed com ICso values in M (SD) of compound (1), cisplatin, and arsenic pounds” and “disclosed compound(s) refer to one ore more trioxide in a panel of different human cancer cell lines compounds having the structure of formulas (I)-(VII), including species therein. Cell line Malignancy Complex 1 Cisplatin As2O3 30 Not all of the depicted components illustrated or A2780 Ovarian cisplatin 20.3 + 4.0 3.1 - 11 17.1 - 1.5 described may be required. In addition, some implementa sensitive tions and embodiments may include additional components. A2780CF Ovarian cisplatin 21.4 + 1.8 47.3 + 2.1 21.6 + 1.4 resistant Variations in the arrangement and type of the components MDA-MB- Triple negative 9.5 O.1 22.3 - 28 11.9 2.3 may be made without departing from the spirit or scope of 231 breast cancer 35 the claims as set forth herein. Additional, different or fewer (mCherry) components may be provided and components may be RPMI 8226 Multiple 4.5 - 1.0 1.9 O.1 7.1 O2 myeloma combined. Alternatively or in addition, a component may be HTC-116 Colon 1.6 + 0.4 5.5 - 1.3 9.4 O.9 implemented by several components. U-87 Glioblastoma O.37 - 0.11 9.6 O.8 1.6 2.9 The above description illustrates the invention by way of 40 example and not by way of limitation. This description clearly enables one skilled in the art to make and use the Example 23 invention, and describes several embodiments, adaptations, variations, alternatives and uses of the invention, including Orthotopic Cancer Model Prophetic Example what is presently believed to be the best mode of carrying 45 out the invention. Additionally, it is to be understood that the The anticancer activity of these compounds will be tested invention is not limited in its application to the details of for efficacy in triple negative breast cancer, ovarian cisplatin construction and the arrangement of components set forth in resistant cancer, multiple myeloma, glioblastoma and colon the following description or illustrated in the drawings. The cancer, as well as in other cancers in mouse Xenograft invention is capable of other embodiments and of being models. The mice will be randomized into 4 treatment 50 practiced or carried out in various ways. Also, it will be groups (8 mice per group) as follows: PBS alone, arseno understood that the phraseology and terminology used platin compounds (4 mg/kg), ASO (4 mg/kg) and cisplatin herein is for the purpose of description and should not be (4 mg/kg). Stock Solutions of arsenic trioxide will be pre regarded as limiting. pared by dissolving solid arsenic trioxide in 5 MNaOH, and Having described aspects of the invention in detail, it will the stock solution will be diluted with PBS and pH adjusted 55 be apparent that modifications and variations are possible to be 7.4. Stock solutions of arsenoplatins and cisplatin will without departing from the scope of aspects of the invention be diluted with PBS. Each group will be treated twice weekly for three weeks by i.p. injections in the case of solid as defined in the appended claims. As various changes could tumors. Tumors will be measured with digital calipers and be made in the above constructions, products, and methods tumor volume will be calculated using the equation VTu without departing from the scope of aspects of the invention, 60 it is intended that all matter contained in the above descrip mor-(w2x1xt)/6. Mice will be weighted twice weekly. tion and shown in the accompanying drawings shall be DEFINITIONS interpreted as illustrative and not in a limiting sense. The technical effects and technical problems in the speci When introducing elements of aspects of the embodi fication are exemplary and are not limiting. It should be ments, the articles “a,” “an,” “the, and “said are intended 65 noted that the embodiments described in the specification to mean that there are one or more of the elements. The may have other technical effects and can solve other tech terms "comprising.”99 “including,”& and “having are intended nical problems. US 9,499,574 B2 43 44 What is claimed is: thioester, imino, C-Co alkyl, C-Coalkenyl, alkynyl, 1. A compound having the structure of formula (I) alkoxyl, amino, amidyl, immino, Sulfonyl, Sulfoxyl, phosphoryl, phosphoryl ester, glycosyl, aryl, C-Cls (I) cycloalkyl, heteroaryl, and C-Cls heterocycloalkyl, i 5 wherein RandR are independently selected from hydro Z-M-4 gen, C-Co alkyl, C-Co alkenyl, alkynyl, alkoxyl, \,: amino, amidyl, immino, Sulfonyl, Sulfoxyl, phosphoryl, V ?y phosphoryl ester, glycosyl, aryl, C-C cycloalkyl, 2 -j-Z heteroaryl and C-Cls heterocycloalkyl; Yi Y2 10 wherein the Lewis base can be halogen, N-bonded cyano, nitroso, nitroxyl, N-bonded C-C alkyl, alkenyl, wherein M is Pt, Pd or Ni; Q is As, Sb or Bi; Z is N: Z N-bonded C-C cycloalkyl, N-bonded heteroalkyl, is O or S; L and L are independently C(O), C R' or N-bonded C-C heteroalkenyl, N-bonded aryl, C R; X is a Lewis base;Y' and Y are independently N-bonded heteroaryl, N-bonded C-Cls heterocy selected from -OR. -SR and - SR", wherein R' 15 cloalkyl, - NRR, N-bonded ligand, S-bonded cyano, and R are independently selected from hydrogen, sulfonyl, sulfoxyl, thiol, S-bonded thioether, S-bonded halogen, cyano, keto, ester, ether, thiol, thioether, thio- thioester, S-bonded C-C alkyl, C-C alkenyl, ester, imino, C-Co alkyl, C-Co alkenyl, alkynyl, S-bonded C-C cycloalkyl, S-bonded C-Co het alkoxyl, amino, amidyl, immino, Sulfonyl, Sulfoxyl, eroalkyl, S-bonded C-C heteroalkenyl, S-bonded phosphoryl, phosphoryl ester, glycosyl, aryl, C-Cls aryl, S-bonded heteroaryl, S-bonded C-Cls heterocy cycloalkyl, heteroaryl, and C-C, heterocycloalkyl: 20 cloalkyl, - SR, S-bonded ligand, O-bonded C-Co wherein Rand Rare independently selected from hydro- cycloalkyl, O-bonded C-C heteroalkyl, O-bonded gen, C-C alkyl, C-C alkenyl, alkynyl, alkoxyl, C-C heteroalkenyl, O-bonded aryl, O-bonded het amino, amidyl, immino, Sulfonyl, Sulfoxyl, phosphoryl, eroaryl, O-bonded C-Cls heterocycloalkyl, -OR. phosphoryl ester, glycosyl, aryl, C-C cycloalkyl, O-bonded carboxylato, O-bonded polycarboxylato, heteroaryl, and C-Cls heterocycloalkyl; 25 O-bonded ligand, or P-bonded phosphine having for wherein the Lewis base can be halogen, N-bonded cyano, mula P(R), (R), where x is 0, 1, 2 or 3: nitroso, nitroxyl, N-bonded C-Co alkyl, alkenyl, wherein RandR are independently selected from hydro N-bonded C-C cycloalkyl, N-bonded heteroalkyl, gen, O. —CO.R. —COR. C-Clo alkyl, C-Clo alk N-bonded C-C heteroalkenyl, N-bonded aryl, enyl, aryl, heteroaryl, C-C cycloalkyl and C-Cls N-bonded heteroaryl, N-bonded C-Cls heterocy- 30 heterocycloalkyl: cloalkyl, - NRR, N-bonded ligand, S-bonded cyano, wherein R and R are independently selected from Sulfonyl, Sulfoxyl, thiol, S-bonded thioether, S-bonded C-C alkyl, C-C alkenyl, aryl, heteroaryl, C-Cls thioester, S-bonded C-Co alkyl, C-Co alkenyl, cycloalkyl and C-Cls heterocycloalkyl; and S-bonded C-C cycloalkyl, S-bonded C-Co het wherein R is selected from hydrogen, hydroxyl, halogen, eroalkyl, S-bonded C-Co heteroalkenyl, S-bonded amine, thiol, ether, C-Co alkyl, C-Coalkenyl, aryl, aryl, S-bonded heteroaryl, S-bonded C-Cls heterocy 35 heteroaryl, C-C cycloalkyl and C-Cls heterocy cloalkyl, - SR, S-bonded ligand, O-bonded C-Co cloalkyl. cycloalkyl, O-bonded C-C heteroalkyl, O-bonded 3. The compound of claim 1, wherein the compound has C-Co heteroalkenyl, O-bonded aryl, O-bonded het the structure of formula (VI): eroaryl, O-bonded C-Cls heterocycloalkyl, —OR, O-bonded carboxylato, O-bonded polycarboxylato, 9 (VI) O-bonded ligand, or P-bonded phosphine having for- H H mula P(R)-(R), where x is 0, 1, 2 or 3: Y-M-N wherein Rand Rare independently selected from hydro- A. t gen, O. —CO.R. —COR, C-Clio alkyl, C-Clo alk enyl, aryl, heteroaryl, C-C cycloalkyl and C-C 45 heterocycloalkyl: wherein R7 and R are independently selected from C-Co alkyl, C-Coalkenyl, aryl, heteroaryl, C-Cls wherein M is Pt, Pd or Ni; Q is As or Sb: X is a Lewis cycloalkyl and C-Cls heterocycloalkyl, and base; wherein R is selected from hydrogen, hydroxyl, halogen, so wherein R' and Rare independently selected from hydro amine, thiol, ether, C-Coalkyl, C-C alkenyl, aryl, gen, halogen, cyano, keto, ester, ether, thiol, thioether, heteroaryl, C-C cycloalkyl and C-Cls heterocy thioester, imino, C-Co alkyl, C-Coalkenyl, alkynyl, cloalkyl. alkoxyl, amino, amidyl, immino, Sulfonyl, Sulfoxyl, 2. The compound of claim 1, wherein the compound has phosphoryl, phosphoryl ester, glycosyl, aryl, C-Cls the structure of formula (II): 55 cycloalkyl, heteroaryl, and C-Cls heterocycloalkyl, (II) and H H wherein the Lewis base can be halogen, N-bonded cyano, N- -N1 nitroso, nitroxyl, N-bonded C-Co alkyl, alkenyl, A. M N-bonded C-C cycloalkyl, N-bonded heteroalkyl, R- C-R*, 60 N-bonded C-C heteroalkenyl, N-bonded aryl, 8––6 N-bonded heteroaryl, N-bonded C-Cls heterocy 3 /V 4 cloalkyl, - NRR, N-bonded ligand, S-bonded cyano, RO OR sulfonyl, sulfoxyl, thiol, S-bonded thioether, S-bonded wherein M is Pt, Pd or Ni; Q is As, Sb or Bi: X is a Lewis thioester, S-bonded C-Co alkyl, C-Co alkenyl, base; 65 S-bonded C-C cycloalkyl, S-bonded C-Co het wherein R' and Rare independently selected from hydro eroalkyl, S-bonded C-Co heteroalkenyl, S-bonded gen, halogen, cyano, keto, ester, ether, thiol, thioether, aryl, S-bonded heteroaryl, S-bonded C-Cls heterocy US 9,499,574 B2 45 46 cloalkyl, - SR, S-bonded ligand, O-bonded C-Co -continued cycloalkyl, O-bonded C-C heteroalkyl, O-bonded (17) C-Co heteroalkenyl, O-bonded aryl, O-bonded het eroaryl, O-bonded C-Cls heterocycloalkyl, —OR, O-bonded carboxylato, O-bonded polycarboxylato, O-bonded ligand, or P-bonded phosphine having for mula P(R)-(R), where x is 0, 1, 2 or 3: wherein Rand Rare independently selected from hydro gen, O. —CO.R. —COR, C-Clio alkyl, C-Clo alk enyl, aryl, heteroaryl, C-C cycloalkyl and C-Cls (18) heterocycloalkyl: 10 wherein R and R are independently selected from C-C alkyl, C-C alkenyl, aryl, heteroaryl, C-Cls cycloalkyl and C-Cls heterocycloalkyl, and wherein R is selected from hydrogen, hydroxyl, halogen, amine, thiol, ether, C-Co alkyl, C-Coalkenyl, aryl, 15 heteroaryl, C-C cycloalkyl and C-Cls heterocy cloalkyl. 4. The compound of claim 3, wherein the compound is selected from the group consisting of compounds (11)-(19): (19)

(11) V / N-P-N 25 A. y HC-C, C-CH3, V O-As-O M. V. HC-C HO OH O-Sb-O (12) 30 A V H C H HO OH \ / N-P-N CHCH A. y CHCH 5. The compound of claim 1, wherein the compound has 3 2- V (W -- 2 3. the structure of formula (VII): O-As-O 35 / V HO OH (13) (VII) H C H V / N-N-N A. y 40 HC-C, C-CH3, V. O-As-O / V HO OH (14) 45 H C H V / wherein X is a Lewis base; N-N-N wherein R' and Rare independently selected from hydro CHCH A. y CHCH gen, halogen, cyano, keto, ester, ether, thiol, thioether, 3 2- V ?y -- 2 3. thioester, imino, C-C alkyl, C-C alkenyl, alkynyl, OHAS- O 50 alkoxyl, amino, amidyl, immino, Sulfonyl, Sulfoxyl, / V HO OH phosphoryl, phosphoryl ester, glycosyl, aryl, C-Cls (15) cycloalkyl, heteroaryl, and C-Cls heterocycloalkyl, H C H V / and N-Pt-N wherein the Lewis base can be halogen, N-bonded cyano, A. y 55 nitroso, nitroxyl, N-bonded C-Co alkyl, C-Co alk HC-C, C-CH3, V enyl, N-bonded C-C cycloalkyl, N-bonded het O-Sb-O eroalkyl, N-bonded C-Co heteroalkenyl, N-bonded / V HO OH aryl, N-bonded heteroaryl, N-bonded C-Cls heterocy (16) cloalkyl, - NRR, N-bonded ligand, S-bonded cyano, H C H 60 sulfonyl, sulfoxyl, thiol, S-bonded thioether, S-bonded \ | / thioester, S-bonded C-C alkyl, C-C alkenyl, N-P-N A. y S-bonded C-C cycloalkyl, S-bonded C-Co het HC-C, C-CH3, eroalkyl, S-bonded C-C heteroalkenyl, S-bonded N ?/ OHSb - O aryl, S-bonded heteroaryl, S-bonded C-Cls heterocy / V 65 cloalkyl, - SR, S-bonded ligand, O-bonded C-Co HO OH cycloalkyl, O-bonded C-C heteroalkyl, O-bonded C-Co heteroalkenyl, O-bonded aryl, O-bonded het US 9,499,574 B2 47 48 eroaryl, O-bonded C-Cls heterocycloalkyl, -OR. -continued O-bonded carboxylato, O-bonded polycarboxylato, (7) H SCN H. O-bonded ligand, or P-bonded phosphine having for V / mula P(R)-(R), where x is 0, 1, 2 or 3: N-Pt-N wherein RandR are independently selected from hydro w C-CH2CH3, gen, O. —CO.R. —COR, C-Clio alkyl, C-Clo alk enyl, aryl, heteroaryl, C-C cycloalkyl and C-Cls heterocycloalkyl: (8) wherein R and R are independently selected from 10 C-Co alkyl, C-Coalkenyl, aryl, heteroaryl, C-Cls cycloalkyl and C-Cls heterocycloalkyl; and wherein R is selected from hydrogen, hydroxyl, halogen, w C-CH2CH3, amine, thiol, ether, C-Co alkyl, C-Coalkenyl, aryl, heteroaryl, C-C cycloalkyl and C-Cls heterocy 15 cloalkyl. (9) 6. The compound of claim 5, wherein the compound is selected from the group consisting of compounds (1)-(10):

(1) V / N-P-N A. y 25 HC-C, C-CH3, V O-As- O (10) M V HO OH (2) H C H 30 V / N-Pt-N CHCH A. y CHCH 3 2- N.. ?y u- 2 3. HO OCH OHAS- O / V 35 HO OH 7. A pharmaceutical composition comprising: (3) H I H an excipient, and V / a compound having the structure of formula (I) N-Pt-N A. y 40 HC-C, C-CH3, N ?y (I) OHAS- O HO OH (4) 45 H\ I /H N-Pt-N CHCH A. y CHCH 3 2- N. ?/ u- 2 3. O-As- O / V 50 wherein M is Pt, Pd or Ni; Q is As, Sb or Bi; Z' is N: Z. HO OH is O or S; L and L are independently C(O), C R' or (5) H SCN H. C R; X is a Lewis base;Y' and Y are independently \ | / selected from OR. -SR and - SR", wherein R' N-Pt-N and R are independently selected from hydrogen, A. y 55 HC-C, C-CH3, halogen, cyano, keto, ester, ether, thiol, thioether, thio V. O-As-O ester, imino, C-C alkyl, C-C alkenyl, alkynyl, M. V. alkoxyl, amino, amidyl, immino, Sulfonyl, Sulfoxyl, HO OH phosphoryl, phosphoryl ester, glycosyl, aryl, C-Cls (6) cycloalkyl, heteroaryl, and C-Cls heterocycloalkyl: H NCS H 60 \ | / wherein RandR are independently selected from hydro N-Pt-N gen, C-Co alkyl, C-Co alkenyl, alkynyl, alkoxyl, A. y HC-C, C-CH3, amino, amidyl, immino, Sulfonyl, Sulfoxyl, phosphoryl, N W phosphoryl ester, glycosyl, aryl, C-C cycloalkyl, O-As-O / V 65 heteroaryl, and C-Cls heterocycloalkyl; HO OH wherein the Lewis base can be halogen, N-bonded cyano, nitroso, nitroxyl, N-bonded C-Co alkyl, alkenyl, US 9,499,574 B2 49 50 N-bonded C-C cycloalkyl, N-bonded heteroalkyl, eroaryl, O-bonded C-Cls heterocycloalkyl, -OR. N-bonded heteroalkenyl, N-bonded aryl, N-bonded O-bonded carboxylato, O-bonded polycarboxylato, heteroaryl, N-bonded C-Cls heterocycloalkyl, O-bonded ligand, or P-bonded phosphine having for —NR'R'', N-bonded ligand, S-bonded cyano, sulfonyl, mula P(R)-(R), where x is 0, 1, 2 or 3: sulfoxyl, thiol, S-bonded thioether, S-bonded thioester, wherein Rand Rare independently selected from hydro S-bonded C-C alkyl, C-Co alkenyl, S-bonded gen, O. —CO.R. C1-Cio alkyl, C-Clo alkenyl, aryl, Cs-Co cycloalkyl, S-bonded C-Co heteroalkyl, heteroaryl, C-C cycloalkyl and C-Cls heterocy S-bonded C-C heteroalkenyl, S-bonded aryl, cloalkyl: S-bonded heteroaryl, S-bonded C-Cls heterocy wherein R and R are independently selected from cloalkyl, - SR, S-bonded ligand, O-bonded C-Co 10 C-Co alkyl, C-Coalkenyl, aryl, heteroaryl, C-Cls cycloalkyl, O-bonded C-C heteroalkyl, O-bonded cycloalkyl and C-Cls heterocycloalkyl; and C-Co heteroalkenyl, O-bonded aryl, O-bonded het wherein R is selected from hydrogen, hydroxyl, halogen, eroaryl, O-bonded C-Cls heterocycloalkyl, —OR, amine, thiol, ether, C-Co alkyl, C-Coalkenyl, aryl, O-bonded carboxylato, O-bonded polycarboxylato, heteroaryl, C-C cycloalkyl and C-Cls heterocy O-bonded ligand, or P-bonded phosphine having for 15 cloalkyl. mula P(R)-(R), where x is 0, 1, 2 or 3: wherein Rand Rare independently selected from hydro 9. The pharmaceutical composition of claim 7, wherein gen, O. —CO.R. —COR, C-Clio alkyl, C-Clo alk the compound has the structure of formula (VI): enyl, aryl, heteroaryl, C-C cycloalkyl and C-Cls heterocycloalkyl: wherein R and R are independently selected from (VI) C-C alkyl, C-C alkenyl, aryl, heteroaryl, C-Cls cycloalkyl and C-Cls heterocycloalkyl, and wherein R is selected from hydrogen, hydroxyl, halogen, amine, thiol, ether, C-Co alkyl, C-Coalkenyl, aryl, 25 heteroaryl, C-C cycloalkyl and C-Cls heterocy cloalkyl. 8. The pharmaceutical composition of claim 7, wherein the compound has the structure of formula (II): 30 wherein M is Pt, Pd or Ni; Q is As or Sb: X is a Lewis base; wherein R' and Rare independently selected from hydro (II) gen, halogen, cyano, keto, ester, ether, thiol, thioether, thioester, imino, C-C alkyl, C-C alkenyl, alkynyl, 35 alkoxyl, amino, amidyl, immino, Sulfonyl, Sulfoxyl, phosphoryl, phosphoryl ester, glycosyl, aryl, C-Cls cycloalkyl, heteroaryl, and C-Cls heterocycloalkyl: and wherein the Lewis base can be halogen, N-bonded cyano, 40 nitroso, nitroxyl, N-bonded C-C alkyl, alkenyl, wherein M is Pt, Pd or Ni; Q is As, Sb or Bi: X is a Lewis N-bonded C-C cycloalkyl, N-bonded heteroalkyl, base; N-bonded C-C heteroalkenyl, N-bonded aryl, wherein R' and Rare independently selected from hydro N-bonded heteroaryl, N-bonded C-Cls heterocy gen, halogen, cyano, keto, ester, ether, thiol, thioether, cloalkyl, - NRR, N-bonded ligand, S-bonded cyano, thioester, imino, C-Co alkyl, C-Coalkenyl, alkynyl, 45 sulfonyl, sulfoxyl, thiol, S-bonded thioether, S-bonded alkoxyl, amino, amidyl, immino, Sulfonyl, Sulfoxyl, thioester, S-bonded C-Co alkyl, C-Co alkenyl, phosphoryl, phosphoryl ester, glycosyl, aryl, C-Cls S-bonded C-C cycloalkyl, S-bonded C-C het cycloalkyl, heteroaryl, and C-Cls heterocycloalkyl, eroalkyl, S-bonded C-Co heteroalkenyl, S-bonded wherein Rand Rare independently selected from hydro aryl, S-bonded heteroaryl, S-bonded C-C heterocy gen, C-Co alkyl, C-Co alkenyl, alkynyl, alkoxyl, 50 cloalkyl, - SR, S-bonded ligand, O-bonded C-Co amino, amidyl, immino, Sulfonyl, Sulfoxyl, phosphoryl, cycloalkyl, O-bonded C-C heteroalkyl, O-bonded phosphoryl ester, glycosyl, aryl, C-C cycloalkyl, C-Co heteroalkenyl, O-bonded aryl, O-bonded het heteroaryl and C-Cls heterocycloalkyl, eroaryl, O-bonded C-Cls heterocycloalkyl, -OR. wherein the Lewis base can be halogen, N-bonded cyano, O-bonded carboxylato, O-bonded polycarboxylato, nitroso, nitroxyl, N-bonded C-Co alkyl, alkenyl, 55 O-bonded ligand, or P-bonded phosphine having for N-bonded C-C cycloalkyl, N-bonded heteroalkyl, mula P(R), (R), where x is 0, 1, 2 or 3: N-bonded C-C heteroalkenyl, N-bonded aryl, wherein RandR are independently selected from hydro N-bonded heteroaryl, N-bonded C-Cls heterocy gen, O. —CO.R. C1-Cio alkyl, C-Clo alkenyl, aryl, cloalkyl, - NRR, N-bonded ligand, S-bonded cyano, heteroaryl, C-C cycloalkyl and C-Cls heterocy sulfonyl, sulfoxyl, thiol, S-bonded thioether, S-bonded 60 cloalkyl: thioester, S-bonded C-C alkyl, C-C alkenyl, wherein R7 and R are independently selected from S-bonded C-C cycloalkyl, S-bonded C-Co het C-Co alkyl, C-Coalkenyl, aryl, heteroaryl, C-Cls eroalkyl, S-bonded C-C heteroalkenyl, S-bonded cycloalkyl and C-Cls heterocycloalkyl; and aryl, S-bonded heteroaryl, S-bonded C-Cls heterocy wherein R is selected from hydrogen, hydroxyl, halogen, cloalkyl, - SR, S-bonded ligand, O-bonded C-Co 65 amine, thiol, ether, C-Co alkyl, C-Coalkenyl, aryl, cycloalkyl, O-bonded C-C heteroalkyl, O-bonded heteroaryl, C-C cycloalkyl and C-Cls heterocy C-Co heteroalkenyl, O-bonded aryl, O-bonded het cloalkyl. US 9,499,574 B2 51 52 10. The pharmaceutical composition of claim 9, wherein -continued the compound is selected from the group consisting of (19) compounds (11)-(19): (11) is Y k \ "'y.A. yy-ll F --yi ; 7. 10 HC-C C-CH3. HO OH V ?/ O-Sb - O (12) A V Fl Ji HO OH N-P-N A. M 15 CHCH- V C-CH2CH3, f-As-d M R 11. The pharmaceutical composition of claim 7, wherein HO OH the compound has the structure of formula (VII): C (13) 2O HV /H N-N-N (VII) HC-C,A. y C-CH3, HQ i H Y-k-6 25 --y Ho Yon R!-c. -R, (14) Y-k-6 Fl Ji HOM. V.OH N-N-N A. M 30 CHCH- V -CHCH, wherein X is a Lewis base; o-A-0 wherein RandR are independently selected from hydro HO Yon gen, halogen, cyano, keto, ester, ether, thiol, thioether, C (15) 35 thioester, imino, C-C alkyl, C-C alkenyl, alkynyl, F. Ji alkoxyl, amino, amidyl, immino, Sulfonyl, Sulfoxyl, --, phosphoryl, phosphoryl ester, glycosyl, aryl, C-Cls Hic- 3-CH, cycloalkyl, heteroaryl, and C-Cls heterocycloalkyl, f-Sb-d and Ho Yon 40 wherein the Lewis base can be halogen, N-bonded cyano, (16) 1itroso, s nitroxyl,y1, N-bonded C-C- O alkyl,3 C-C-Y-O alk H C H enyl, N-bonded C-C cycloalkyl, N-bonded het \-a-N eroalkyl, N-bonded C-Co heteroalkenyl, N-bonded H c-c^ -CH 45 aryl, N-bonded heteroaryl, N-bonded C-Cls heterocy 3 V. f7 3. cloalkyl, - NRR, N-bonded ligand, S-bonded cyano, O-Sb-O / V SU lfonvil.y1, Sulfoxvl,y1, thiol.s S-bonded thioether.s S-bonded HO OH thioester, S-bonded C-Co alkyl, C-Co alkenyl, C (17) S-bonded C-C cycloalkyl, S-bonded C-Co het F. Ji 50 eroalkyl, S-bonded C-Co heteroalkenyl, S-bonded -- aryl, S-bonded heteroaryl, S-bonded C-C heterocy HC-C C-CH3, cloalkyl, - SR, S-bonded ligand, O-bonded C-C N8-it-6 y cycloalkyl, O-bonded C-C heteroalkyl, O-bonded3. O 55 C-Co heteroalkenyl, O-bonded aryl, O-bonded het HO OH (18) eroaryl,O O-bonded C-Clss heterocycloalkyl,s -OR. 21 O-bonded carboxylato, O-bonded polycarboxylato, O-bonded ligand, or P-bonded phosphine having for N 60 lala P(R7), (R(K), Whereh X 1Sis 0,U, 1, 2Z or 33: F. Ji wherein RandR are independently selected from hydro N-Pt-N gen, O. —CO.R. —COR. C-Clo alkyl, C-Clo alk Hic- 3-ch. and enyl, aryl, heteroaryl, C-C cycloalkyl and C-Cls heterocycloalkyl: ot-o 65 wherein R and R are independently selected from HO OH C-Co alkyl, C-Coalkenyl, aryl, heteroaryl, C-Cls cycloalkyl and C-Cls heterocycloalkyl; and